237Pu experiments with the plaice Pleuronectes platessa

Some experiments have been made on the metabolism of ²³⁷Pu — a high specific activity, gamma-emitting isotope of plutonium — by the plaice Pleuronectes platessa L. Very little of the isotope was incorporated into fish after 2 months exposure to labelled water. The oral retention from a variety of labelled foods was also very poor, and apart from the gut itself no incorporation of the isotope could, in fact, be demonstrated. Injected ²³⁷Pu was eliminated very slowly, and some evidence was obtained for an inverse relationship between the rate of ²³⁷Pu loss and the rate of growth of the fish. Growing fish incorporated a relatively larger fraction of the ²³⁷Pu body burden into skeletal material, and this was attained at the expense of the isotopic content of the liver. Very little ²³⁷Pu was incorporated into muscle.     

237Pu experiments with the thornback ray Raja clavata

Some experiments have been made on the metabolism of ²³⁷Pu — a high specific activity, gamma-emitting isotope of plutonium — by the thornback ray Raja clavata L. Unlike previous experiments with the plaice (Pleuronectes platessa L.) it was found that the thornback ray absorbed plutonium across the gut wall, as readily observed by analysis of the liver. The liver accumulated up to 0.2% of the ²³⁷Pu given in a single labelled meal after a period of 12 days. Direct injection of the isotope into the muscle of thornback rays resulted in extremely slow rates of elimination from the fish, the highest internal concentrations being attained by the spleen. The largest fractions of the whole-body burden, however, were those of liver and skeleton.     

Transuranic nuclides in plaice (Pleuronectes platessa) from the North-Eastern Irish Sea

The concentrations of a number of -emitting nuclides, ²³⁸Pu, ^239/240Pu, ²⁴¹Am, ²⁴²Cm and ^243/244Cm have been determined in the organs of plaice, Pleuronectes platessa L., caught in the vicinity of a nuclear fuel reprocessing plant. Fish were taken for analysis every 3 months for a period of 2 years. During this period, the highest concentrations of plutonium and americium nuclides were found in the kidney and the lowest in muscle. In all of the organs analysed, the concentrations of americium were greater than those of plutonium: they were also greater than plutonium when related to the rates of discharge of these two elements and gave higher concentration factors over samples of filtered shore-line sea water taken from the area.     

Biokinetics of plutonium,americium and californium in the marine isopod Cirolana borealis,with observations on its feeding and molting behavior

In a series of laboratory experiments carried out during 1982, the biokinetics of ²³⁷Pu (IV), ²³⁷Pu (V), ²⁴¹Am (III), and ²⁵²Cf (III) were studied in the benthic marine isopod Cirolana borealis Lilljeb. Following a 3 wk exposure to labelled seawater, isopods reached non-equilibrium concentration factors of 52±15, 54±12, 176±14, and 185±53 for Pu (IV), Pu (V), Am and Cf, respectively. Uptake of these radionuclides occurred to a great extent by surface adsorption, with the major fraction of the accumulated radionuclides being associated with exoskeleton and lesser percentages located in gut, digestive gland, muscle and haemolymph. The pattern of radiotracer depuration was in all cases biphasic, with the long-lived retention compartment retaining the largest fraction of whole-body radioactivity. Biological half-lives for radionuclide turnover in the long-lived compartment were 60±3 d, 87±5 d, 261±12 d and 288±60 d for Pu (IV), Pu (V), Am and Cf, respectively. Examination of radionuclide distribution among tissues following longterm depuration showed that the radionuclides were eliminated from internal tissues more rapidly than from the exoskeleton. Uptake and depuration studies indicated that the biokinetics of americium and californium were similar to each other, but that they differed significantly from those of oxidized and reduced plutonium. In general, these experimental results support previous reports that americium and californium are more bioavailable than plutonium in the marine waters. The fraction americium and californium assimilated into tissue from food was very low and did not exceed 5%. Unassimilated labelled food was often retained in the gut for more than 2 mo and released only infrequently in a loosely bound condition. In the event of contamination, this particular feeding-digestion strategy in highly mobile, scavenging isopods could act as a potential biological mechanism for the widespread dispersion of radioactive wastes.     

Studies on the nutrition of marine flatfish. The effect of dietary protein content on certain cell components and enzymes in the liver of Pleuronectes platessa

In order to examine the effects of artificial diets at the tissue level, plaice (Pleuronectes platessa L.) were fed either high (50%) or low (20%) protein diets for several weeks and the total quantities of deoxyribonucleic acid (DNA), ribonucleic acid (RNA), protein and phospholipid in their livers were measured. Alanine amino-transferase, aspartate aminotransferase, glutamate dehydrogenase and glucose-6-phosphatase activities in the livers were also measured. Similar measurements were made on the livers of wild plaice. The total amount of DNA in the livers of both cultured and wild plaice, of standard weight, did not differ significantly, irrespective of dietary history. In fish of standard weight, total hepatic RNA, protein and phospholipid levels were highest in the cultured fish fed high protein diet and lowest in the wild fish. These differences were related to dietary protein supply and environmental temperature. The total organ levels of the liver enzymes in plaice of equal weight were not significantly different in the cultured fish fed high and low protein diets. Wild-fish livers contained a smaller total amount of alanine aminotransferase, aspartate aminotransferase and glucose-6-phosphatase activity than did those of cultured fish. Total liver glutamate dehydrogenase activity was not significantly different in any of the fish examined. It is suggested that kinetic and allosteric factors may be more important in controlling protein metabolism and amino acid cat abolism in fish than total enzyme level.     

Single and combined effects of heavy metals and hormones on lysosomes of haemolymph cells from the mussel Mytilus galloprovincialis

Effects of heavy metals on lysosomes were studied in living cells from the mussel (Mytilus galloprovincialis Lam.). Haemolymph cells were obtained from the mussel adductor muscle, stained with neutral red (NR), and analysed by digital imaging to evaluate NR retention times within lysosomes. Exposure to Hg²⁺, Cd²⁺ and Cu²⁺ induced a reduction of NR retention time, indicating lysosomal membrane destabilisation. The intensity of these effects was correlated with the metal affinity for sulfhydryls. In contrast, Zn²⁺ showed no effect on lysosomes. Moreover, 200 μM Zn²⁺ protected lysosomes against the effects of Cd²⁺ and Cu²⁺, but not against Hg²⁺. Cell loading with the fluorescent pH probe Lyso Sensor followed by digital imaging showed a rise of lysosomal pH induced by Cd²⁺ and Hg²⁺, while Zn²⁺ prevented the effect of Cd²⁺ and also partially that of Hg²⁺. The different protective effect of Zn²⁺ against Hg²⁺ suggests a dual action of Hg²⁺ on lysosomes, possibly involving both membrane destabilisation and proton pump inhibition. Cell exposure to 17 β-estradiol also caused a reduction of NR retention time, which was synergistic to that of Hg²⁺. This suggests a common pathway between metals and hormone, possibly involving Ca²⁺ signaling. Received: 17 November 1999 / Accepted: 29 June 2000     

Studies on the nutrition of marine flatfish. The effect of dietary fatty acids on the tissue fatty-acids of the plaice Pleuronectes platessa

The ratios of polyunsaturated fatty acids in the lipids of hatchery-reared plaice [Pleuronectes platessa (L.)] maintained on a diet containing corn oil and freeze-dried cod muscle accurately reflect ratios of polyunsaturated fatty acids in the diet. Such plaice have much greater levels of triglycerides in both their livers and extrahepatic tissues than wild plaice of comparable size; the total amount of 22:6 3 acid in triglycerides of the two groups is, however, very similar. Transfer of experimental plaice to a fat-free diet caused a marked depletion of triglycerides from both livers and extrahepatic tissues with almost all fatty acids being reduced in amount. Addition of 12:0 and 14:0 ethyl esters (4% of each) to the fat-free diet caused some accumulation of triglycerides in extrahepatic tissues. Further addition of 18:2 6 and 18:3 3 acids (both at levels of 0.4%) to the diet caused an accumulation of triglycerides in the liver. These triglycerides had increased levels of saturated and monounsaturated fatty acids up to C 18 but the levels of higher polyunsaturated fatty acids were not inereased. There was relatively little change in the fatty acids of phospholipids throughout these experimental dietary regimes. The levels of phospholipids were constant and very similar to those of wild plaice. The results indicate: (1) plaice fed a diet relatively low in 22:6 3 acid may accumulate relatively large amounts of triglyceride in order to store adequate amounts of 22:6 3 acid; (2) plaice are capable of synthesising saturated and mono-unsaturated fatty acids up to C 18 in the presence of dietary 12:0 and 14:0 acids; (3) 18:2 6 and 18:3 3 acids in the diet do not undergo chain elongation and further desaturation in plaice with the dietary regimes we used.     

Nitrogen balance in marine fish larvae: influence of developmental stage and prey density

The utilization and fate of nitrogen in larvae of plaice (Pleuronectes platessa), blenny (Blennius pavo) and herring (Clupea harengus), from the stage of first-feeding to metamorphosis, was examined under laboratory conditions. Rates of ammonia excretion, primary amine defaecation, and growth in terms of protein-nitrogen were monitored throughout larval life. Data were used to calculate daily ration, the coefficient of nitrogen utilization (absorption efficiency), and gross and net growth efficiencies. The developmental pattern of nitrogen balance was similar for plaice and blenny larvae. These species showed increasing growth efficiency (k₁: 55 to 80%) with decreasing weight-specific waste nitrogen losses with age. Absorption efficiencies. were high (83 to 98%) in plaice and blenny larvae, and tended to increase with development in the former species. Ration relative to body weight decreased with growth in both species. Herring larval development, although at a slower rate than blenny and plaice, appeared normal up to 33 d, after which high mortality occurred. Absorption efficiency in this species tended to decline (83 to 43%) with age, until metabolic costs exceeded the absorbed ration and growth ceased. Artemia sp. nauplii proved a suitable food source for the rearing of plaice and blenny larvae, but this diet may have long-term toxicity or deficiency effects on herring. Availability and density of food affected nitrogen balance in the larvae of all three species. Feeding stimulated the output of wastes in excretion and defaecation by a factor of up to ten times the 12-h non-feeding basal rates. Waste nitrogen output reached a peak some 2 to 3 h after commencement of feeding and returned slowly to the baseline in 5 to 10 h after cessation of feeding. There was an asymptotic increase in ration, ammonia output and growth of larvae as prey density increased. Ration saturated at a higher prey density (>4 prey ml^-1) than either growth or excretion rate (1 prey ml^-1). Thus the efficiency with which food is absorbed and utilized for growth must eventually decline in response to high prey density. The idea that larval fish are adapted to maximize ingestion and growth rate, rather than optimize growth efficiency and thus to respond to prey occurring in either low density or in occasional patches, is supported by these results.     

Effects of oil dispersants on the developing embryos of marine fish

The effects of oil dispersants BP1002¹, Finasol ESK and Corexit 7664 on the development of herring, Clupea harengus L., plaice, Pleuronectes platessa L. and sole, Solea solea (L.) have been studied. Treatment of developing embryos for 100 h with BP1002 and Finasol ESK gave rise to abnormalities in cell division and differentiation, to reductions in heart-rate, eye pigmentation, growth rate and hatching success above concentrations of 10 parts/10⁶. Larvae with abnormal flexures of the spine, which prevented them from feeding successfully, resulted from exposure to 5 parts/10⁶. Treatment of the embryos with dispersants throughout the period from fertilisation to hatching (15 to 20 days) produced similar abnormalities at slightly lower concentrations. Corexit did not produce any demonstrable deleterious effects on embryos exposed to concentrations up to 5000 parts/10⁶, the highest tested.     

Studies on digestion and the fine structure of digestive caeca in Eurydice pulchra (Crustacea: Isopoda)

In the carnivorous isopod Eurydice pulchra Leach the mid-gut caeca have a pH of 5.6 and produce a strong acid proteinase, a lipase and a carbohydrase. Electron microscope studies of the digestive caeca of starved and fed animals show that secretion is merocrine and may originate from a complex involving the basal cell membrane and associated mitochondria, producing lysosomes. Absorbed materials, in fed animals, appear to take the form of neutral lipid bodies, glycogen, and proteinaceous crystals, the latter forming within the golgi complex. Digestion of both lipid and protein inclusions was observed to occur intra-cellularly by fusion with free lysosomes. Secretion and absorption may occur in the same cell, and the scheme for the sequence of epithelial cell stages is proposed.     

Ultrastructural responses and oxidative stress induced by cypermethrin in the liver of Labeo rohita

The present study was conducted to establish the relationship between selected oxidative stress parameters and ultrastructural responses in liver tissue of Labeo rohita fingerlings exposed to cypermethrin. Fish were exposed to lethal (4.0 μg L^?1) and sublethal (0.4 μg L^?1) concentrations of cypermethrin for a period of 24, 48, 72 and 96 h for acute studies and 1, 5, 10 and 15 days for subacute studies, respectively. Results showed increased catalase (CAT) and protease activity, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), protein carbonyls and free amino acid (FAA) levels at both concentrations. This suggests participation of free-radical-induced oxidative cell injury in mediating the hepatotoxicity of cypermethrin. In corroboration of this, ultrastructural lesions witnessed a reduction in the number of cell organelles, swollen, vacuolated and condensed mitochondria, dilated rough endoplasmic reticulum, and reduced numbers of smooth enodplasmic reticulum, peroxisomes and lysosomes at the lethal (4.0 μg L^?1) concentration. At the sublethal (0.4 μg L^?1) concentration, cytoplasmic vacuolation, condensed, vacuolated and swollen mitochondria, dilated rough endoplasmic reticulum and an absence of hepatocyte microvilli were prominent. Ultrastructural changes were exhibited as subcellular responses due to the imbalance in cellular oxidative status by means of oxidative damage.     

Etude préliminaire,in situ,de la distribution du plutonium dans différents tissus et organes de Cancer pagurus (Crustacea: Decapoda) et de Pleuronectes platessa (Pisces: Pleuronectidae)

Plutonium distribution was preliminary investigated in tissues and organs of the crab Cancer pagurus and the plaice Pleuronectes platessa collected inshore near the La Hague fuel reprocessing plant (France). There is an important transfer of plutonium from sea water to the gills and exoskeleton of the crab, these organs representing a large surface adsorption. In the plaice, only the gut strongly accumulates plutonium, indicating contamination by feeding. The edible parts of these two marine species, particularly the flesh, do not constitute, for man, an important source of contamination by environmental plutonium.     

Experimental studies on the effects of copper on a marine food chain

Effects of copper sulphate on a marine food chain were investigated in large tanks at Loch Ewe on the west coast of Scotland. The food chain consisted of phytoplankton, the bivalve Tellina tenuis da Costa, and O-group plaice (Pleuronectes platessa L). During initial months after settlement, T. tenuis siphons are an important food for juvenile plaice, which feed by grazing the siphon tips; these subsequently regenerate. Copper dose rates of 10, 30 and 100 μg Cu/1 were investigated; the distribution of added copper was monitored and its metabolic effects were determined. Copper levels in water, sand, in algae on tank walls, in T. tenuis shell and flesh, and in plaice muscle and viscera were measured by atomic absorption spectrophotometry. Copper accumulated in sand, T. tenuis flesh and shell, and plaice viscera. Accumulations were dose-dependent and in no case was a plateau concentration reached. Most of the added dose was taken up by the sand; accumulations in T. tenuis shell were small — less than twice background in all cases. High levels were found in T. tenuis flesh, where concentrations after 100 days were 270, 470 and 1100 μg Cu/g dry flesh for dose concentrations of 10, 30 and 100 μg Cu/1, respectively. The mean control concentration was less than 50 μg Cu/g dry flesh. There was no accumulation of copper in plaice muscle. Visceral accumulations after 100 days were 71, 147 and 567 μg Cu/g dry flesh for dose concentrations of 10, 30 and 100 μg Cu/1, respectively. The mean control level was 30 μg Cu/g dry flesh. The effect of copper on phytoplankton metabolism was investigated by measuring plant pigment levels and rates of primary photosynthetic fixation of C¹⁴-labelled bicarbonate. All dose concentrations reduced both the standing crop and the rate of photosynthesis per unit of chlorophyll a. The effects of copper on growth and condition of T. tenuis and P. platessa were investigated. All dose concentrations adversely affected T. tenuis condition (dry flesh weight for standard individual). The effect was most marked during deposition of winter reserves. In the absence of plaice predation, there was a decrease in the mean siphon weight for T. tenuis exposed to 30 and 100 μg Cu/1. For plaice, all dose concentrations of copper resulted in reduced growth, but there was no significant change in condition or biochemical composition. The ash weight of fish exposed to copper was significantly higher than for controls.     

Morphological and histological changes during the growth and starvation of herring and plaice larvae

Reared herring (Clupea harengus L.) and plaice (Pleuronectes platessa L.) were examined for morphological and histological changes during growth and starvation. The growth rate of herring larvae of 0.22 mm/day was less than that reported for wild stock, but this difference was attributed to survival of runts in laboratory. Larval plaice had a growth rate of 0.16 mm/day. The relative condition factor (antilogarithm of intercept of length-weight line) was used to assess condition throughout the larval stages. Starvation resulted in a progressive collapse of the larval body, especially of the ventral body surface around the pectoral girdle of both species (assessed by the pectoral angle) and of the spacing between the organs of the head in herring. There was a breakdown of the herring gut with decreases in epithelial cell height and catabolism of the connective tissue coat and a marked reduction in the transverse sectional area of the plaice liver. The changes in the pectoral angle in both herring and plaice and the eye height to head height ratio in herring should be useful to fishery biologists for assessing nutritional condition, even on board ship.     

Ultrastructural and radiotracer studies of pore function in foraminifera

Clusters of mitochondria have been observed close to the inner end of the pores in Nonionella stella, Globobulimina pacifica, Buliminella tenuata, Bolivina argentea, Loxostomum pseudobeyrichi, Cassidulinoides cornuta and Bolivina cf. subexcavata, pointing to a pore function of oxygen-uptake. The relationship between mitochondria and pores is most distinct in Buliminella tenuata, Bolivina argentea and L. pseudobeyrichi collected from a deep-water, low-oxygen environment. Amphistegina lobifera Larsen, 1976 readily took up ¹⁴CO₂ through its pores, indicating a pore function related to the photosynthetic activities of its symbionts. ¹⁴C-labelled glucose did not pass through the pores in traceable amounts during a 2 h exposure time.     

Essential and nonessential elements in the red-crowned crane Grus japonensis of Zhalong Wetland,northeastern China

The concentrations of four essential (Ca, Mg, Zn, and Cu) and two nonessential elements (Pb and Cd) in feathers and kidneys, livers, gut walls, and muscles of eight carcasses of migratory red-crowned cranes (Grus japonensis) from Zhalong Wetland, northeastern China, were examined. The concentrations of Cd in the feathers were between 0.4 mg kg^?1 dry weight (dw) and 3.1 mg kg^?1 dw, in the livers between 0.4 and 4.4 mg kg^?1 dw, the maximum of which exceeded a level considered to be environmental exposure risk (i.e., 3 mg kg^?1 dw in the liver or kidney). High Pb levels (0.4–3.2 mg kg^?1 dw, with an average of 1.8 mg kg^?1) were also detected in livers, which exceeded a level considered toxicosis in birds (1.7 mg kg^?1 dw). Pb and Cd had the highest scores in principal component analysis. Relatively high Pb and Cd concentrations in the migratory cranes were thought to be associated with their habitat and prey.     

Some effects of food density on the growth and behaviour of plaice larvae

The effect of food density on the growth, survival, and swimming activity of plaice larvae (Pleuronectes platessa L.) was investigated under controlled laboratory conditions. Suboptimal food concentrations decrease the growth of the height of the body musculature relative to length, and increase the amount of time spent searching for food. Older larvae are able to withstand much longer periods without food than young larvae. On the basis of these experiments, it is suggested that food limitation in a larval plaice population is likely to result in a concave mortality curve.     

Metallothioneins and lysosomes in metal toxicity and accumulation in marine mussels: the effect of cadmium in the presence and absence of phenanthrene

It has been confirmed that metallothioneins play an important role in the accumulation of cadmium (Cd) in the digestive gland cells of mussels (Mytilus galloprovincialis Lam.). The content of Cd in the tissue of mussels exposed for 9 d to the metal (estimated dosage of 180 g Cd mussel^-1 d^-1) was 66.2 ppm. This value is about the same as the metal content found in the digestive gland of Cd-exposed mussels kept in clean water for a recovery period of 28 d. At the end of the recovery period, however, the Cd bound to thionein had increased by approximately 250%. Our data demonstrate that the stability of lysosomes, a biological parameter adopted as a cellular stress index, is extremely low in mussels exposed to Cd for 9 d, but returns to control values in the digestive gland cells of mussels allowed to recover for 28 d in uncontaminated sea water. At this point most of the Cd present in the cytosol is bound to thionein. These data demonstrate the importance of metallothionein induction in the reduction of the cytotoxic effects exerted by high levels of Cd accumulation. The results of tests designed to clarify the reasons for the long biological half-life of Cd demonstrated that, in the digestive gland of mussels, the lysosomes are not able to eliminate Cd either bound to insoluble thionein polymers or to lipid peroxidation products such as lysosomal lipofuscin, both of which are apparently involved in the elimination of copper. The absence of these two mechanisms of metal sequestration and elimination via excretion of residual bodies (tertiary lysosomes) is in agreement with the persistence of cadmium in the digestive gland of mussels. Finally, the results also demonstrate that simultaneous exposure of mussels to Cd and phenanthrene, an established lysosomal membrane destabilizer, did not significantly alter the accumulation of Cd or the kinetics of the metal in mussels.     

The food of four species of pleuronectiform larvae in the eastern English Channel and southern North Sea

An examination was made of the stomach contents of the larvae of the plaice Pleuronectes platessa Linnaeus, 1758; the flounder Platichthys flesus (Linnaeus, 1758), the dab Limanda limanda (Linnaeus, 1758), and the sole Solea solea (Linnaeus, 1758) collected in the eastern English Channel and in the Southern Bight during the winter and spring of 1971. These 4 species of flat fish have distinct diets, and competition for food between them is largely avoided. Plaice larvae fed almost exclusively on Oikopleura dioica; flounder larvae also ate O. dioica, but in addition a wide range of planktonic organisms including phytoplankton, polychaete larvae, lamellibranch larvae, and copepod nauplii. Dab larvae fed mainly on the nauplii and copepodite stages of a variety of copepods, but particularly of Temora longicornis. Some T. longicornis copepodites and polychaete larvae were eaten by sole larvae, but the principal prey of these was lamellibranch larvae. The larvae of all the species began to feed in the yolk-sac stage; the initial food of all except plaice consisted of dino-flagellates, followed by tintinnids and copepod nauplii. Feeding began at dawn and the number of feeding fish and the number of food organisms in their stomachs increased throughout the day to a maximum near sunset. There were no consistent differences between the two areas in the diets of any of the species.     

Ultrastructure and anaerobic metabolism of mitochondria in the marine oligochaete Tubificoides benedii : effects of hypoxia and sulfide

It has often been suggested that ultrastructural properties of mitochondria are correlated with oxygen and sulfide levels from the environment, although careful analyses of this question are rare. In this study the ultrastructure and distribution of mitochondria in Tubificoides benedii, a marine oligochaete from sulfide-rich sediments, were investigated after a series of oxic, hypoxic and hypoxic–sulfidic (200 μM H₂S) incubations up to 24 h. Succinate, one of the key endproducts of an anaerobic metabolism, was used as an indicator of mitochondrial anaerobiosis. Consistent differences in mitochondrial ultrastructure were not observed in any of the incubations, even after 24 h. Stereological parameters of mitochondria (volume density, surface density of the outer mitochondrial membrane, and specific surface) in epidermal and intestinal tissues of T. benedii were not affected by hypoxia or sulfide either. On the other hand, succinate concentrations increased significantly within 24 h under hypoxic and hypoxic–sulfidic conditions. Thus, experimental hypoxia and sulfide clearly caused mitochondrial anaerobiosis without affecting ultrastructure or distribution of mitochondria in T. benedii. Distinct differences in ultrastructural and stereological parameters were common between different tissues and between individuals, showing that different forms of mitochondria can occur within one species. Our results imply that a mitochondrial ultrastructure specific to thiobiotic animals does not appear to exist. Received: 4 August 1996 / Accepted: 20 September 1996