Some effects of prolonged starvation on the metabolism of the red and white myotomal muscles of the plaice Pleuronectes platessa

The effect of starvation on glycogen, lipid, water content and muscle proteins of red and white muscles in the plaice Pleuronectes platessa L. has been investigated. The two muscle systems behave differently in their response to food shortage, glycogen and protein depletion being more marked in the white muscle. Little change occurred in the constituents of the red muscle, except lipid. Starvation resulted in an increase in water content in the white, but not in the red muscle. The differential response of these two musole systems to starvation is discussed. It is suggested that the small amount of red muscle in the body is spared from the effects of starvation because it is needed for continuous activity during swimming at all speeds.     

Tidal and circadian activity rhythms in juvenile plaice,Pleuronectes platessa

Young plaice (Pleuronectes platessa L.), caught in the intertidal zone, exhibit a short-lived tidal rhythm when their activity is recorded in darkness in the laboratory. This tidal rhythm rapidly changes to one of circadian frequency both in darkness and in light-dark cycles. It is considered that the basic rhythm is circadian in nature, but that it can be entrained to keep in phase with the tides by some, as yet unknown, Zeitgeber present under tidal conditions.     

Further 237Pu experiments with the plaice Pleuronectes platessa: Subcellular distribution of plutonium in the liver

The subcellular distribution of ²³⁷Pu in the livers of the plaice Pleuronectes platessa has been studied by differential and isopycnic gradient centrifugation. It has been shown that the ²³⁷Pu is particularly accumulated in pellets sedimenting out at 3000 and 6000 g which, by deduction from marker enzyme tests, contain a mixture of mitochondria and lysosomes. Separation of lysosomes from mitochondria was achieved by using a non-lytic detergent injected into the fish 3 d prior to removal of the livers; it was found that ²³⁷Pu was associated with the lysosomal fraction. In this respect, the metabolism of plutonium of the plaice resembles that of the rat.     

Influence of temperature on muscle-fibre development in larvae of the herringClupea harengus

The development of swimming (myotomal) muscles was studied in herring larvae (Clupea harengus L.) caught in the Firth of Clyde, Scotland, in spring 1990 and reared at either 5, 10 or 15°C. Two muscle-fibre types can be distinguished in the myotomes of herring larvae using ultrastructural criteria. A single layer of small-diameter muscle fibres, packed with mitochondria, is present beneath the entire surface of the skin (superficial muscle fibres). The remaining bulk of the muscle is composed of larger diameter fibres (inner muscle fibres) containing significantly more myofibrils than the superficial fibres. In 1 d-old larvae, the number of inner muscle fibres in myotomes immediately posterior to the yolk-sac was 311±41 at 15°C, 257±22 at 10°C and 187±22 at 5°C (mean±SD,n=6). The average diameter of inner muscle fibres increased with decreasing temperature, so that the total cross-sectional area of muscle was similar at each temperature. After 6 to 7 d, the number of muscle fibres had significantly increased at 15°C (383±25), but not at 10°C (281±32) or 5°C (192±17). In contrast, the average cross-sectional area of inner muscle fibres had increased by 19% at 15°C, 34% at 10°C, and 26% at 5°C. Temperature also influenced the relative proportions and spatial distributions of muscle-fibre organelles. For example, in 1 d-old larvae, the fraction of muscle-fibre volume (volume density) occupied by mitochondria in the superficial fibres was significantly higher at 15°C (46.0%) than at either 5°C (37.6%) or 10°C (38.8%). In the inner muscle fibres, the volume density of mitochondria was 26.1% at 15°C, 20.5% at 10°C and 15.9% at 5°C, whereas the volume density of myofibrils was similar at the three temperatures (33 to 38%). Typically, inner muscle fibres from 10°C larvae, but not from 5 or 15°C larvae contained a large central mitochondrion.     

Development of cell types in the pituitary of Anguilla anguilla,Pleuronectes platessa and Limanda limanda

Pituitary cell types have been studied by application of specific histochemical methods. Identification of cell types in the fishes Anguilla anguilla, Pleuronectes platessa and Limanda limanda at different seasons and maturity stages is based on both tinctorial and structural characteristics of different cell types; their granulation, state of vacuolation, condition of the nucleus and presence or absence of the nucleolus. Gonads from fish caught monthly were also prepared for histological examination to determine their maturity stage and sex. Female A. anguilla at maturity stages II and III, and yellow in all external morphological characters, were used. A general cytohistological study of the pituitary gland during the annual cycle has been carried out with the aim of discovering (a) the causes of the longterm growth, and (b) the pituitary's physiological control of the fishes' metamorphosis. Variations in number and activity of the cell types to be found in the meso adenohypophysis have been investigated by cell counts of representative medial sagittal sections of the pituitary during different months. somatotrophs show short-term fluctuations, but gonadotrophs show an alternating cycle of increase and decrease. Chromophobes do not show much change in their number during the annual life-cycle. Basophils are found in all 3 lobes of the adenohypophysis and exhibit very little structural difference, so that a common function for all 3 cell types may be assumed. Observations on variations in concentration of neurosecretory material have been made; peak accumulation of neuroscretory material occurs between August and October and then remains almost constant up to December. This periodic increase suggests that neurosecretory substances have an influence upon the animal's metabolic activities. Studies in relation to the maturation cycle carried out on monthly samples of male and female P. platessa and L. limanda regarding the percentage, relative numbers and state of activity of chromophobes, somatotrophs and gonadotrophs produced results which appear to agree with the functions suggested for each cell group; their variation in number and activity keeps pace with corresponding maturity phases. Somatotrophs predominate in all maturity stages in both sexes. Chromophobes exhibit a cyclic increase and decrease; few are present in ripening gonads, many in spawning individuals. Gonadotrophs show a relative increase in number during gonadal ripening. Active and inactive gonadotrophs are identifiable at all maturity stages. The eel pituitary size is independent of both length and weight; however, for P. platessa pituitary size is related to weight, which is (in part) dependent upon the maturity stage. Implications of variations in relative proportions of cell types are discussed with regard to present knowledge of the pituitary gland.     

Studies on the cytolytic effects of seastar (Marthasterias glacialis) saponins and synthetic surfactants in the plaice Pleuronectes platessa

The toxic effects of Marthasterias glacialis (L.) saponins and synthetic surface-active agents were studied using the plaice Pleuronectes platessa L. as test animal. Solutions of M. glacialis saponins caused considerable, although non-uniform, damage to the gill epithelium, death resulting from a disturbance in the levels of oxygen, carbon dioxide, and sodium ions in the tissues and blood. Histological examination showed that solutions of the synthetic surfactants caused less damage to the gill epithelium than did the M. glacialis saponins. There was a linear relationship between the logarithm of the median haemolytic concentration and the logarithm of the median lethal concentration for the synthetic surfactants. Haemolytic inhibition experiments suggest that the M. glacialis saponins damage cell membranes by interacting with membrane cholesterol.     

Orientation of plaice (Pleuronectes platessa) in the open sea: evidence for the use of external directional clues

We studied the orientation of plaice (Pleuronectes platessa L.) in the open sea (Southern Bight of the North Sea) during May and June of 1979 and 1980, using continuous, on-line data from 6 fish equipped with transponding acoustic compass tags. During the day, fish swimming in midwater did not maintain their heading. At night, the same fish maintained their heading in a way that was consistent with active orientation to some external directional clue, but inconsistent with the use of an inertial guidance system. We believe this to be the first analysis to demonstrate the use of external guidance clues by fish in the open sea. Because fish were able to maintain a consistent heading in midwater by night, in the apparent absence of both visual and tactile clues, the external reference is likely to be geophysical. Possible clues include the electrical field generated by the flow of sea water through the geomagnetic field and the earth's magnetic field itself.     

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.     

Studies on the nutrition of marine flatfish. Growth of the plaice Pleuronectes platessa on diets containing proteins derived from plants and other sources

The growth of 0-group Pleuronectes platessa L. on diets containing plant proteins was examined. Diets were made acceptable to the fish by including in them cod flesh which had been predigested with proteolytic enzymes, as well as a proportion of air-dried cod meal. A leaf-protein concentrate was fed at 6 levels ranging from 0 to 63 % of the total dietary protein; B.P. protein concentrate (Toprina) and soyabean meal were examined at a single level — about 45 % of the dietary protein. The remainder of the protein in these diets was animal protein, cod meal or cod flesh. Analysis of the weight gains after a 12 week feeding period and of the growth curves showed that, as the proportion of leaf protein in the diet increased, the growth rate diminished, the effect becoming more pronounced when leaf protein reached 40 % or more of the dietary protein. Growth on the diet containing B.P. protein concentrate was superior to that on the diet containing soyabean meal and was comparable with diets containing low levels of leaf protein. The protein efficiency ratios (PER= g live weight gain/g protein fed) for the different diets showed similar trends to the growth rates. Diets containing 40 % or less of leaf protein and the B.P. protein diet had PERs of 1.4 to 1.5. The soyabean meal diet and diets containing more than 40 % of the protein as leaf protein had PERs of 1.0 or less. The results are considered in the context of production diets for flatfish.     

Sublethal impairment of respiratory control in plaice (Pleuronectes platessa) larvae induced by UV-B radiation, determined using a novel biocybernetical approach

The increasing amount of UV-B radiation (280 to 320 nm) reaching the earth's surface could endanger transparent fish and zooplankton in upper water layers. Previous studies on marine fish eggs and larvae focused mainly on the effects of high, lethal doses of ultraviolet irradiation. Now a sensitive test for accumulated, sublethal impairments of physiological performance has been established. In developing fish larvae (Pleuronectes platessa L.), the respiratory control responses to hypoxia were evaluated by combining a biocybernetical approach and a novel optical test system. Before testing respiratory control behavior, the larvae were irradiated with light containing UV-B (test individuals) or free from it (control individuals). Differences in the physiological reactions of these larvae were detected by biocybernetical analysis (linear systems analysis) of respiratory control. The analysis was based on the evaluation of changes in ventilatory rate caused by sinusoidally modulated partial pressures of ambient oxygen. The ventilatory movements of the larvae were recorded, analyzed and evaluated by video microscopy and digital image processing. After several days of irradiation with moderate UV-B doses or intensities (8.6 kJ m⁻² d⁻¹, corresponding to an irradiance of 0.24 W m⁻² for 10 h), sublethal impairments of respiratory control could be detected in plaice larvae by this method; both the linearity of respiratory control itself and the parameters of linear control behavior were affected by UV-B. Concerning environmental implications, the results must still be interpreted cautiously, because natural irradiation conditions are more complex than the artificial regimen. Our biocybernetical approach will be useful for further studies on sublethal impairments of physiological functions caused by UV-B in fish and other transparent marine or limnic animal groups. In addition, the method could be used to test the sublethal impact of chemical contaminants. Received: 4 November 1997 / Accepted: 6 May 1998     

Settlement patterns of newly settled plaice (Pleuronectes platessa) in a non-tidal Swedish fjord in relation to larval supply and benthic predators

In demersal fish species with a pelagic larval stage, settlement patterns may be a consequence of variations in larval supply, habitat selection at settlement, and processes acting between the time of settlement and the time of benthic sampling. This study describes temporal (1994-1998) and spatial variation in plaice (Pleuronectes platessa L.) settlement densities in four semi-isolated nursery areas with similar habitat characteristics, in the non-tidal Gullmarsfjord on the west coast of Sweden. Juvenile abundance varied by a factor of ten, both among years and among nursery grounds. For the 3 years when larval sampling was undertaken (1994-1996) and all nursery areas, there was a significant positive relationship between larval supply and juvenile abundance (linear regression: r²=0.45, n=24, P<0.001). On the southern side of the fjord, a significant positive relationship between larval and juvenile abundance was found in one area (r²=0.62, n=6, P<0.05). The absolute mortality rate of plaice after settlement was related to the initial settlement density (r²=0.95, n=20, P<0.001), and to the abundance of predatory shrimps Crangon crangon (r²=0.44, n=20, P<0.01). Plaice otoliths were found in 6% of the shrimp stomachs analysed from an area with high density (13.3 m^-2) of newly settled plaice. The present study suggests that the density of juvenile plaice was limited by larval supply to the nursery grounds. Consistency in the relative abundance of juveniles among nursery grounds between years also suggested that some nursery areas may be in the settlement shadow of others. The irregular nature of the coastline in combination with larval depletion could thereby cause small-scale (10³-10⁴ m) variation in settlement densities of the same order of magnitude as the inter-annual variability in recruitment to individual nursery grounds.     

Microsatellite analysis of plaice (Pleuronectes platessa L.) in the NE Atlantic: weak genetic structuring in a milieu of high gene flow

Many species of marine fish are typified by large population sizes, strong migratory behavior, high fecundity, and pelagic eggs and larvae that are passively transported by ocean currents, all features that tend to increase gene flow, and hence reduce genetic partitioning, among localized populations. The plaice, Pleuronectes platessa, is a commercially important demersal species that exhibits all of these characteristics. We analyzed genetic variation at eight microsatellite loci in samples of spawning adults (N = 348) from the coasts of Ireland, Iceland, and, for the first time, from the Baltic Sea. Significant differentiation was observed between Iceland and Irish and Baltic Sea samples. However, there were no genetic differences between Irish and Baltic Sea samples, which contrast with the significant differentiation reported between Baltic Sea and North Sea/Atlantic populations of other flatfish species. To increase the data set, we carried out a cross-calibration exercise, allowing us to perform a joint analysis of data with an earlier study on adult and juvenile plaice (N = 480) collected over a broad geographic range, using six microsatellite loci in common to the two studies. Significant differentiation was observed between fish collected at the northern (Iceland, Faeroes, Norway) and southern (Bay of Biscay) parts of the species range. In contrast, the results showed little evidence of genetic structuring over much of the continental shelf of Europe. We believe that bathymetric and hydrographic barriers are the major factors shaping genetic structure, while lack of structure over much of the European continental shelf may be explained by a combination of past historical events, population structure, and dynamics of the species.     

Studies on the nutrition of marine flatfish. The effect of oral administration of diethylstilboestrol and cyproheptadine on the growth of Pleuronectes platessa

Two organic compounds, diethylstilboestrol a synthetic oestrogen substitute, and cyproheptadine which acts as an appetite stimulant in some mammals, were added separately at 3 different concentrations to diets fed to plaice (Pleuronectes platessa), and their effect on growth examined. Over a 10 week period, low orally-administered diethylstilboestrol (600 g/kg moist diet) gave rise to increased weight gain, food consumption and food conversion. Higher dietary concentrations of diethylstilboestrol (up to 2400 g/kg diet) were without effect on these parameters. Cyproheptadine did not stimulate appetite, weight gain or food conversion at any of the levels tested.With an Appendix on Statistics by J. A. Pope and A. M. Shanks     

Influence of food concentration,temperature and salinity on the larval development ofBalanus amphitrite

Influence of food concentration (0.5, 1 and 2 x 10⁵ cell ml^–1 ofSkeletonema costatum), temperature (20 and 30°C) and salinity (15, 25 and 35) on the larval development ofBalanus amphitrite (Cirripedia: Thoracica) was examined. The mortality rate at 20°C was lower than at 30°C in general. Increase in food concentration from 0.5 to 1 x 105 cells ml^–1 improved the survival rate, but this was not evident when food concentration was increased to 2 x 105 cells ml^–1. The results indicate that food availability and temperature jointly determine the energy allocation for metamorphic progress. It was observed that the influence of the tested variables varied with instar. At 20 °C the mean duration of the second instar exceeded 3 d and was much longer than other instar durations. The fourth, fifth and sixth instars and the total naupliar period showed that the effect of different salinities at given food concentrations was negligible at 20°C, while at 30°C there was a marked decrease in duration with increasing salinity.     

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.     

Some influences of temperature on the rearing of the grunion Leuresthes tenuis,an atherine fish

Grunion Leuresthes tenuis (Ayres), an atherine fish, were reared, from artificially fertilized eggs to 160 days posthatching, at temperatures from 18.0° to 25.4° C. The rearing was carried out in five 60 l rectangular containers and two hard moulded plastic pools holding 60 and 450 l. The growth rate was highly correlated with temperature at least up to metamorphosis. Thereafter, the effect of temperature on growth, within lethal limits, appeared to be lost or at least masked by other factors. At low densities, after metamorphosis, growth was proportional to the population density. Size and shape of the rearing containers also influenced the growth rate.     

Effect of development rate on the swimming, escape responses, and morphology of yolk-sac stage larval American plaice, Hippoglossoides platessoides

To examine the impact of development rate on swimming performance, escape response, and morphology, yolk-sac larvae of American plaice (Hippoglossoides platessoides, Fabricius) were reared at two temperatures (5 and 10 °C). Videomicroscopy and silhouette collimation videography were used to examine swimming, escape behaviour, and morphology (standard length, finfold area, and yolk-sac area) of individual larvae. Larvae were examined from 0 d post hatch (dph) to 14 dph for the 5 °C treatment group and from 0 to 6 dph for the 10 °C treatment group (3 August to 17 August 1996). Since larvae were not fed, yolk-sac reserves were essentially exhausted by 14 and 6 dph for the 5 and 10 °C treatment groups, respectively. To control for the effect of testing temperature on behaviour, larvae from each temperature treatment were tested at both 5 and 10 °C. Testing temperature had an effect on some swimming parameters but not on escape response. Swimming performance, escape response, and morphology varied with age, while only morphology and escape response varied with development rate. Morphology and swimming performance, and morphology and escape response were found to be correlated as determined by canonical correlation. This study suggests that both types of swimming behaviours should be examined when developing models of the impacts of predation on the early life history of larval fish. Received: 13 September 1999 / Accepted: 21 June 2000     

Influence of temperature and salinity on the uptake,distribution and depuration of mercury,cadmium and lead by the black-lip oyster Saccostrea echinata

Saccostrea echinata (Quoy and Gaimard) were exposed to 10 g 1^-1 of either mercury, cadmium or lead at 30 °C, 36S; 30 °C, 20S; 20°C, 36S and 20°C, 20S for 30 d and were then transferred to clean seawater for a further 30 d to depurate. Specimens were removed at regular intervals during the exposure and depuration periods, dissected into gills, mantle, visceral mass and adductor, and analysed for the appropriate metal by atomic absorption spectroscopy. Mercury was concentrated more than the other metals in all tissues under all conditions. Cadmium uptake was greater than lead in all tissue in the high-temperature experiments, whereas both metals were concentrated to similar extents at low temperature. The gill tissue generally accumulated the greatest amount of all 3 metals, whilst the adductor concentrated the least amount. At both salinities, mercury and cadmium accumulation by all tissues was significantly greater at the higher temperature whereas lead uptake was only marginally increased. The accumulation rates of mercury at high temperature were significantly greater in all tissues at low compared with high salinity, whereas at low temperature, differences were not significant. Accumulation rates of cadmium and lead in the majority of tissues examined were significantly greater in lowsalinity water at both temperatures. In general, lead was lost the most rapidly from oyster tissues, followed by mercury and then cadmium. The residence times for mercury and cadmium differed significantly between tissues, with the gills showing the highest turnover rate. In contrast, residence times for lead were similar between tissues. Losses of all 3 metals from oyster tissues were not obviously influenced by temperature and only mercury losses differed significantly between salinities.     

Laboratory study of predation by Aurelia aurita on larvae of cod,flounder, plaice and herring: development and vulnerability to capture

Capture success of the medusa Aurelia aurita preying on various developmental stages of fish larvae was measured together with larval reactivity and escape speed after being stung. These experiments were conducted in the spring of 1983 with A. aurita medusae collected from Loch Etive, Scotland and laboratory-reared larvae of Gadus morhua L., Platichthys flesus L., Pleuronectes platessa L. and Clupea harengus L. Capture success of the medusae increased with medusa size, but decreased with advancing larval development. Smaller species of larvae were more vulnerable to capture. Larval reactivity to encounters with medusae increased with advancing development, and larger species of larvae were more reactive to encounters. Larval escape swimming speeds also increased with advancing larval development and size. These results indicate that earlier stages of larvae within a species and smaller species of larvae at a given stage are more vulnerable to predation by medusae since they are less reactive to encounters. Apparently they are more susceptible to the effects of neurotoxins. Predation rates on different developmental stages of herring larvae are documented and compared with rates predicted by a predation model. Predictions fell within the range of observed predation rates, but tended to overestimate rates by larger medusae feeding on larger herring larvae. This indicates the possibility of predator satiation and/or behavioural avoidance.     

Abundance,distribution and prey composition of scyphomedusae in the southern North Sea

The annual cycle of abundance and distribution of the scyphozoan medusae Aurelia aurita, Cyanea lamarckii, C. capillata and Chrysaora hysoscella were studied in the southern North Sea in 2004 and 2005. Three different patterns of seasonal occurrence of medusae were distinguished: (1) the early occurring C. lamarckii (February–August), (2) C. capillata and A. aurita (April–August) and (3) the late appearing C. hysoscella (July/August–September). Cyanea lamarckii was the most frequently encountered species in this study; its highest mean abundance was 1.8 ± 2.7 ind. 100 m⁻³. The prey spectra of C. lamarckii, C. capillata and C. hysoscella contained several copepod and other crustacean species and thus make them potential competitors with fish larvae. Medusae in this study also consumed fish eggs and larvae, including clupeids, in all months analysed. Although peak spawning of sprat (Sprattus sprattus) coincides with the maximum abundance of medusae (May–June) the relative low abundance of all medusae species in this study makes jellyfish predation unlikely to be a factor controlling sprat recruitment in the time frame investigated.