RNA/DNA ratio and expression of 18S ribosomal RNA, actin and myosin heavy chain messenger RNAs in starved and fed larval Atlantic cod (Gadus morhua)

Levels of total RNA, total DNA, 18S ribosomal RNA (rRNA), poly(A) messenger RNA (mRNA), and two mRNAs coding for abundant myofibrillar proteins were estimated in laboratory-reared Atlantic cod larvae (Gadusmorhua Linnaeus) under conditions of feeding and starvation. DNA probes specific for cod 18S rRNA, β-actin mRNA and myosin heavy chain mRNA were developed. In two experiments on newly hatched larvae in fed and starved treatments, changes in 18S rRNA and mRNA were similar to changes in total RNA during the first weeks after hatching. RNA levels in fed and starved larvae in both experiments were stable, or increased, over the first 3 d after hatching, and then decreased to minima at 9 d. RNA levels increased after 9 d, with the degree and timing of the increase varying among the individual classes of RNA. Complete mortality of starved larvae in both experiments was observed shortly after 11 d, corresponding to exhaustion of endogenous yolk reserves. Total RNA content, RNA/DNA ratio, 18S rRNA levels, total mRNA pool, and actin and myosin heavy chain mRNA levels showed significant differences in fed and starved first-feeding larvae after yolk exhaustion. In another experiment with 3- to 4-week-old cod larvae, 18S rRNA levels were significantly lower in starved versus fed larvae after 3 d. Total RNA responded to feeding and starvation within a similar time as 18S rRNA and the mRNAs examined. Analysis of bulk nucleic acids using fluorometric dyes was simpler and faster than analysis of individual RNAs using hybridization probes, and provides valuable information on recent growth and condition of individual larvae. However, analysis of specific RNAs can provide information on expression of the corresponding genes and reveal the changes underlying trends seen in bulk RNA. Received: 9 February 1996 / Accepted: 7 June 1999     

Facultative lecithotrophy during larval development of the burrowing shrimp Callianassa tyrrhena (Decapoda: Callianassidae)

Survival, developmental and consumption rate (Artemia nauplii ingested per day) as well as predation efficiency (ingested per available Artemia nauplii) were studied during the larval development of the shallow-water burrowing thalassinid Callianassa tyrrhena (Petagna, 1792), which exhibits an abbreviated type of development with only two zoeal stages and a megalopa. The larvae, hatched from berried females from S. Euboikos Bay (Aegean Sea, Greece), were reared at 10 temperature–food density combinations (19 and 24 °C; 0, 2, 4, 8 and 16 Artemia nauplii d⁻¹). Enhanced starvation resistance was evident: 92 and 58% of starved zoeas I molted to zoea II, while metamorphosis to megalopa was achieved by 76 and 42% of the hatched zoeas at 19 and 24 °C, respectively. The duration of both zoeal stages was affected by temperature, food density and their interaction. Nevertheless, starvation showed different effects at the two temperatures: compared to the fed shrimp, the starved zoeae exhibited accelerated development at 19 °C (8.4 d) but delayed metamorphosis at 24 °C (5.9 d). On the other hand, both zoeal stages were able to consume food at an increased rate as food density and temperature increased. Predation efficiency also increased with temperature, but never exceeded 0.6. Facultative lecithotrophy, more pronounced during the first zoeal stage of C.tyrrhena, can be regarded as an adaptation of a species whose larvae can respond physiologically to the different temperature–food density combinations encountered in the wide geographical range of their natural habitat. Received: 28 February 1998 / Accepted: 21 October 1998     

Effect of unfavorable trophic scenarios on amylase and protease activity of <Emphasis Type="Italic">Nephrops norvegicus</Emphasis> (L.) larvae during their first vertical migration: a laboratory approach

In Portuguese waters, Nephrops norvegicus larvae hatch at 400–800 m depth and need to perform a vertical migration to food-rich shallower waters to find suitable prey. The effect of suboptimal feeding on digestive enzymes activity of N. norvegicus larvae during this early period of their larval life remains unknown. Protease and amylase activities were investigated ex situ using flurometry in laboratory-hatched larvae exposed to different feeding and/or starving scenarios in the 24 h following hatching, the period during which they typically accomplish their upward vertical migration. Amylase activity was very low in comparison with protease activity, indicating that carbohydrates are not a primary energy reserve. Larvae starved for 12 h and subsequently fed displayed no increase in amylase activity, which suggests that feeding may be required before 12 h post-hatch to trigger amylase activity. Protease activity was high under all feeding conditions, and the increase in protease activity under sustained starvation indicated the catabolism of protein reserves. The ability of first-stage N. norvegicus larvae to metabolize protein reserves may play a decisive role for their survival during their first vertical migration, as it enables them to overcome the deleterious effects of short-term starvation and/or suboptimal feeding.     

Influence of cadmium accumulation and dietary status on fatty acid composition in two colour forms of shore crabs, Carcinus maenas

 The effects of cadmium exposure and dietary status on cadmium accumulation, fatty acid (FA) content and profiles were investigated in two colour forms of the shore crab Carcinus maenas. Groups of shore crabs were either starved or fed with blue mussels, Mytilus edulis, during a 40 d exposure period to 2 or 6 μM Cd²⁺ (as CdCl₂). Starved green individuals accumulated more cadmium in haemolymph and hepatopancreas than did red crabs and green crabs fed during the experiments. In the red colour form, no difference in cadmium accumulation was observed between starved and fed individuals. In both colour forms, hepatopancreas contained more FA than gills and muscle. The FAs often present in the largest amounts in the tissues were 16:0, 16:1ω7, 18:1ω7, 18:1ω9, 20:4ω6, 20:5ω3 and 22:6ω3. However, saturated (SAFA) and mono-unsaturated fatty acids (MUFAs) were dominant in hepatopancreas, whereas poly-unsaturated fatty acids (PUFAs) were dominant in gills and muscles. At the beginning of the experiment, the total FA content in the hepatopancreas was 111.6 mg g⁻¹ (dry weight) for red crabs and 78.4 mg g⁻¹ for green shore crabs. During the experiment, however, the FA content decreased in red crabs. This decrease was more pronounced for starved individuals than for fed individuals. Also, the decrease in FA content was more pronounced in crabs exposed to 6 μM cadmium compared to crabs exposed to 2 μM or crabs not exposed to cadmium. No change in FA content was observed in green shore crabs, irrespective of diet and cadmium exposure. For both colour forms, no change in FA content was observed for gills and muscle. In red crabs, a decrease was observed for all FAs in the hepatopancreas. This decrease, however, was more pronounced for SAFAs and MUFAs than for PUFAs, indicating that the metabolism of FAs during starvation and cadmium exposure is selective. The experiments indicate that green colour forms of shore crabs are more tolerant of natural stress such as starvation and anthropogenic stress, e.g. cadmium exposure, than are red colour forms of shore crabs. Received: 23 September 1999 / Accepted: 29 April 2000     

Osmoregulation, nutritional effects and buoyancy of marine larval fish: a bioassay for assessing density changes during the earliest life-history stages

It has been hypothesized that marine fish larvae in the advanced stages of starvation would show increased density (ρ = mass volume⁻¹) from water loss due to osmoregulation failure. Changes in larval buoyancy are currently attributed to swim bladder regulation and protein synthesis or catabolism. Osmoregulation-related changes in density is an alternative mechanism, the importance of which remains untested in the laboratory and the influence of which on vertical distributions is unknown. We provide evidence that loss of osmotic control is a plausible mechanism for increased density of larval cod (Gadus morhua L.). Furthermore, our results show that this mechanism is not restricted to larvae in the advanced stages of starvation. “Relative” larval densities are estimated using a modified density gradient. We use a gravimetric method to separate the effects of nutrition from osmoregulation failure. We assessed the importance of sampling strata on estimates of larval density. Proportional sampling within three depth strata (stratified sample) produced the least biased method for determining the “average” density of a population of larvae in laboratory culture. Larvae sampled from the bottom third of the culture tank were significantly more dense then those sampled from the surface. This was true for larvae of all ages. The average change in density from hatching till death from starvation for larvae sampled in the surface stratum was nominal (Δρ = 5.0 × 10⁻⁴ g cm⁻³), while the change for those sampled from the bottom stratum was large (Δρ = 3.8 × 10⁻³ g cm⁻³). These large density differences suggest that larvae sampled from the bottom stratum were either osmotically stressed or were facultatively changing their density via regulatory pathways. Preliminary observations suggest that vitality is lower amongst those larvae which are sampled near the bottom. The small change in average density of larvae sampled from the surface stratum was due to starvation. The density differences we observed between “osmotically stressed” and “starving” larvae could readily have been misconstrued as differences in feeding and growth experienced by individual larvae. The potential bias of increased density from osmoregulation failure must be considered as a factor in experimental designs developed to assess the effect of fed and starved treatments on buoyancy for larvae of all ages. The simple bioassay we describe may prove useful both as a means of assessing larval condition and as a mechanism for evaluating factors affecting larval vertical distributions in the field. Received: 13 January 1997 / Accepted: 3 February 1997     

Effect of Cd on accumulation and loss of 210Po in the mussel Mytilus edulis

As a result of ²¹⁰Po's previously identified association with sulphur-rich proteins, metallothioneins could have a significant effect on the behaviour and fate of ²¹⁰Po in molluscs. Starved control and cadmium-exposed mussels, Mytilus edulis, were fed ²¹⁰Po-labelled algae (Isochrysis galbana) for 5 d and then allowed to depurate in clean sea water. Cadmium-exposed M. edulis accumulated less ²¹⁰Po in the digestive gland and the remainder of the tissue than control mussels, although this was due to a decrease in tissue weight. More than 40% of ²¹⁰Po was identified as being associated with high molecular weight and heat-treated cytosol proteins in M. edulis. Mussels in a starved state are known to recycle as much as 90% of their amino acids. It is proposed that ²¹⁰Po associated with these and other proteins is recycled, explaining why no significant loss of ²¹⁰Po was observed from the remainder of the tissue in either control or Cd-exposed mussels. Cadmium-induced metallothioneins had no effect on the distribution of ²¹⁰Po in M. edulis; <5% associated with the cytosolic fraction was considered to principally contain metallothioneins. It is suggested that ²¹⁰Po's apparent relationship with metallothioneins is coincidental rather than connected with its role in the regulation of metals. Received: 30 March 1998 / Accepted: 3 August 1998     

Ontogenetic changes in digestive enzyme activity of the spiny lobster,<Emphasis Type="Italic"> Jasus edwardsii</Emphasis> (Decapoda; Palinuridae)

Digestive enzyme profiles of puerulus, post-puerulus, juvenile and adult stages of the spiny lobster Jasus edwardsii Hutton were determined in order to identify ontogenetic changes in digestive capabilities and assess capacity to use dietary components and to exploit diets to meet nutritional requirements. Juvenile and adult lobsters exhibited a diverse range of enzymes, suggesting they can exploit varied diets. Proteolytic activity of trypsin, chymotrypsin and carboxypeptidase A (1.86–3.70 U mg^–1 digestive gland protein) was significantly higher than carbohydrase activity of amylase, -glucosidase, cellulase and chitinase (0.0014–0.38 U mg^–1 digestive gland protein), thus showing that dietary protein was more important than carbohydrate and that the lobster is carnivorous. These conclusions are consistent with other studies that found spiny lobsters to feed predominantly on crabs, bivalves, ophiuroids and sponges. Lipase activity (0.371 U mg^–1 digestive gland protein) was also relatively high, thus showing the importance of dietary lipid. Total activities (units per digestive gland) of every enzyme assayed increased significantly with lobster carapace length. There were few significant ontogenetic trends in specific enzyme activity (U mg^–1 digestive gland protein). Amylase and laminarinase specific activity was significantly higher in small lobsters than large lobsters (regression analyses, respectively, F_(1,23)=9.84, P=0.005; F_(1,11)=19.65, P=0.001), suggesting that carbohydrates including laminarin are more important in the diet of small juveniles. Trypsin, amylase and lipase activities were detected in all non-feeding puerulus stages, suggesting that feeding is not a cue for digestive enzyme production in J. edwardsii. Significant variations in total and specific activities of amylase (F_(1,3)=14.2, P=0.00; F_(1,3)=14.2, P=0.00) and trypsin (F_(1,3)=8.8, P=0.00; F_(1,3)=21.41, P=0.00) and a declining trend in lipase total activity between non-feeding puerulus stages suggests that they may be hydrolysing endogenous energy reserves to sustain their onshore swimming activity prior to settlement. Profiles suggest carbohydrate and lipid are utilised first, followed by protein. Consistently high levels of lipase in all puerulus stages (0.24–0.7 U digestive gland^–1; P>0.05) confirm the importance of lipid as a major energy substrate.Communicated by G.F. Humphrey, Sydney     

Cadmium, zinc and copper accumulation in the squid Illex argentinus from the Southwest Atlantic Ocean

 The levels and burdens of Cd, Zn, Cu and Hg were measured in the dorsal mantle, digestive gland and gonads of the squid Illex argentinus, from the Southwest Atlantic Ocean. Mature and immature individuals of both sexes were analysed. Correlations of heavy metal concentrations and burdens with sex and food habits were studied. The highest metal levels were found, particularly for Cd, in the digestive gland, which attained a concentration of up to 270 μg g⁻¹ (wet weight). Dorsal mantle and gonads exhibited the same order of metal enrichment: Zn>Cu>Cd. Zinc and Cu levels were higher in ovaries than in testes and varied with the stage of maturation. The dorsal mantle exhibited the lowest heavy metal concentrations. Mercury levels were below the detection limit of the method in all the tissues analysed. Received: 7 September 1999 / Accepted: 9 March 2000     

Effect of temperature on statolith growth of the European squid Loligo vulgaris during early life

Over the past decade, statolith interpretation has resulted in a major advance in our knowledge of squid population-dynamics, but the way in which environmental conditions affect the statolith increment-deposition ratio remains virtually unknown. The object of the present study was to determine the effect of temperature on this process, using tetracycline marks to validate statolith growth in Loligo vulgaris Lamarck, 1798 under rearing conditions equivalent to severe winter (11 °C) and summer (19 °C) temperature regimes. Tetracycline marking was performed every 10 d (at 10, 20, 30, 40, 50 and 60 d of age). The newly hatched squid paralarvae were slightly smaller in summer than those hatched in winter. Survival rates were similar in both cultures, but growth rates (wet mass) of summer squids were double those in winter. At hatching, statoliths were already longer in the summer squids, and growth rates were 2% d⁻¹ as opposed to 0.9% d⁻¹ for winter statoliths. For the dorsal dome area of the statolith, where more increment counts were made, statolith growth was of 3.25 μm d⁻¹ in summer, and daily increment deposition was confirmed in 87% of the statoliths. The slow growth of statoliths at winter temperatures yielded a mean growth of 1.1 μm d⁻¹– insufficient to discern the increments using light microscopy. Subsequent SEM observation enabled only 21% of the winter statoliths to be read; these also indicated a deposition rate of one increment d⁻¹. Since the life span of L. vulgaris is ≃1 yr, squids will experience at least one winter during their life cycle, and this might be visible on the statolith. Received: 28 June 1999 / Accepted: 20 December 1999     

Condition of larval and juvenile red drum (Sciaenops ocellatus ) from estuarine nursery habitats

RNA:DNA ratios of larval and juvenile red drum (Sciaenops ocellatus) collected from nursery habitats in the Aransas Estuary, Texas, in 1994 were quantified using a highly sensitive ethidium-bromide fluorometric technique. RNA:DNA ratios of wild red drum were evaluated by comparing individual values to a linear regression model derived for starved laboratory-reared red drum. Wild red drum were in relatively good condition with <5% of the RNA:DNA ratios within or below the 95% prediction interval of 4 to 5 d starved red drum. A multiple-regression model explained 54% of the variability in the RNA:DNA ratio of wild red drum, and identified length and water temperature (midday) as significant factors. RNA:DNA ratios increased with fish length [≃1.1 mm⁻¹, over the size range investigated (5␣to 20 mm)]. The effect of temperature on the RNA: DNA ratio was assessed on different sampling trips, and ratios increased with increasing temperature. Abundance of larval and juvenile red drum in the Aransas Estuary varied as a function of both habitat (shoal grass Halodule wrightii, turtle grass Thalassia testudinum) and site (Aransas Bay, Redfish Bay); however, no differences in RNA:DNA ratios were detected between habitats or between sites. It is postulated that the nutritional condition of newly settled red drum from the Aransas Estuary in 1994 was relatively high, and that starvation was of minor importance. Received: 19 August 1996 / Accepted: 23 August 1996     

Zooplankton responses to hypoxia: behavioral patterns and survival of three species of calanoid copepods

Seasonally recurrent and persistent hypoxic events in semi-enclosed coastal waters are characterized by bottom-water dissolved oxygen (d.o.) concentrations of < 2.0 ml l⁻¹. Shifts in the distribution patterns of zooplankters in association with these events have been documented, but the mechanisms responsible for these shifts have not been investigated. This study assessed interspecific differences in responses to hypoxia by several species of calanoid copepods common off Turkey Point, Florida, USA: Labidocera aestiva (Wheeler) (a summer/fall species), Acartia tonsa (Dana) (a ubiquitous year-round species), and Centropages hamatus (Lilljeborg) (a winter/spring species). Under conditions of moderate to severe hypoxia 24-h survival experiments were conducted for adults and nauplii of these species from August 1994 to October 1995. Experiments on adults used a flow-through system to maintain constant d.o. concentrations. Adults of A. tonsa showed no decline in survival with d.o. as low as 1.0 ml l⁻¹, sharp declines in survival at d.o. = 0.9 to 0.6 ml l⁻¹, and 100% mortality with d.o. = 0.5 ml l⁻¹. Adults of L. aestiva and C. hamatus were more sensitive to oxygen depletion: both species experienced significant decreases in survival for d.o. = 1.0 ml l⁻¹. Nauplii of L. aestiva and A. tonsa showed no significant mortality with d.o. = 1.1 to 1.5 ml␣l⁻¹ and d.o. = 0.24 to 0.5 ml l⁻¹, respectively. In addition, experiments investigating behavioral avoidance of moderate to severe hypoxia were carried out for adults of all three species. None of the three species effectively avoided either severely hypoxic (d.o. < 0.5 ml l⁻¹) or moderately hypoxic (d.o. ≈ 1.0 ml l⁻¹) bottom layers in stratified columns. These results suggest that in␣nearshore areas where development of zones of d.o. < 1.0 ml l⁻¹ may be sudden, widespread, or unpredictable, patterns of reduced copepod abundance in bottom waters may be due primarily to mortality rather than avoidance. Received: 31 August 1996 / Accepted: 24 September 1996     

Effects of municipal piers on the growth of juvenile fishes in the Hudson River estuary: a study across a pier edge

The growth rates of two fish species, the winter flounder Pseudopleuronectes americanus (Walbaum) (19.3 to 42.6 mm total length, TL) and the tautog Tautogaonitis (Linnaeus) (23.9 to 55.9 mm TL), were used to evaluate habitat quality under and around municipal piers in the Hudson River estuary, USA. Growth rates were measured in a series of 10 d field caging-experiments conducted at two large piers in the summers of 1996 and 1997. Cages (0.64 m²) were deployed along␣transects that stretched from underneath the piers to beyond them, encompassing the pier edge (the transitional zone between the pier interior and the outside). Growth in weight (G _w ) was determined at five locations along the transect, 40 m beneath the pier, 20 m beneath the pier, at the pier edge, 20 m beyond the pier edge, and 40 m beyond. Under piers, mean growth rates of winter flounder and tautogs were negative (xˉG _W = −0.02 d⁻¹), and rates were comparable to laboratory-starved control fishes (xˉG _W = −0.02 d⁻¹). In contrast, mean growth rates at pier edges and in open waters beyond piers were generally positive (xˉG _W ranged from −0.001 to +0.05 d⁻¹), with growth at pier edges often being more variable and less rapid than at open-water sites. Analyses of stomach contents upon retrieval of caged fishes revealed that dry weights of food were generally higher among fishes caged at open-water stations (xˉ range = 0.02 to 0.72 mg dry wt) than at pier-edge (xˉ range = 0.01 to 0.54 mg) or under-pier (xˉ range = 0.03 to 0.11 mg) stations, although it was apparent that benthic prey were available at all stations on the transect. Our results indicate poor feeding conditions among fishes caged under piers, and suboptimal foraging among fishes caged at pier edges. Inadequate growth rates can lead to higher rates of mortality, and, based on these and other earlier experiments, we conclude that under-pier environments are poor-quality habitats for some species of juvenile fishes. Received: 12 March 1998 / Accepted: 9 November 1998     

Lipid classes and their content of n-3 highly unsaturated fatty acids (HUFA) in Artemia franciscana after hatching, HUFA-enrichment and subsequent starvation

The distribution of n-3 highly unsaturated fatty acids (HUFA) over the major neutral and polar lipid classes was determined for two predominant types of live food used in the larviculture of marine fish and shrimp, i.e. freshly hatched and HUFA-enriched Artemia, and compared with data reported in the literature for wild copepods, representing the natural diet of these larvae. Lipid class composition and their content of n-3 HUFA, particularly eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), were assessed in freshly hatched, HUFA-enriched and subsequently starved Artemia franciscana. The n-3 HUFA enrichment was based on feeding Artemia a lipid emulsion in which either fatty acid ethyl esters (EE, diluted with olive oil) or triacylglycerol (TAG) provided a level of 30% n-3 HUFA. Enrichment of Artemia with either type of the lipid emulsions resulted in an increase of total lipid content from 20.0 to 28.2–28.7% of dry matter mainly due to the accumulation of neutral lipid, primarily TAG (from 82 to 158 mg g⁻¹ dry wt in freshly hatched and 24-h enriched Artemia). Enriched brine shrimp utilized up to 27–30% of their TAG content during 72 h of starvation at 12 °C. The absolute tissue concentrations of polar lipids remained constant at 71 to 79 mg g⁻¹ dry wt throughout the enrichment and subsequent starvation. The level of n-3 HUFA increased drastically during enrichment from 6.3% of total fatty acids (8.2 mg g⁻¹ dry wt) in freshly hatched nauplii to between 20.4 and 21.8% (40.4 to 43.2 mg g⁻¹ dry wt) in 24-h enriched Artemia and was not significantly affected by the source of n-3 HUFA. During starvation, 18:0, 20:4n-6 and 20:5n-3 were retained, whereas 18:4n-3, 22:5n-3 and 22:6n-3 were specifically catabolized. The major polar lipids, phosphatidylethanolamine (PE) and phosphatidylcholine (PC), of freshly hatched Artemia showed very low levels of DHA (<0.1% of total fatty acids) and carried about 45% of the total EPA present. Enrichment with either of the emulsions resulted in an increase of the neutral lipid fraction which concentrated >64% of the EPA and >91% of the total DHA present. This is in sharp contrast with the high levels of n-3 HUFA, in particular DHA, in the polar lipid fraction reported for wild copepods. The contrasting distribution of DHA in the neutral and polar lipid fractions of enriched brine shrimp compared to the natural diet may influence the efficacy of this essential fatty acid for marine fish larvae in aquaculture systems. Received: 10 June 1997 / Accepted: 8 August 1997     

Effects of monospecific and mixed-algae diets on survival, development and fatty acid composition of penaeid prawn (Penaeus spp.) larvae

Four species of microalgae (Chaetoceros muelleri, Tetraselmis suecica, Tahitian Isochrysis sp. (T-iso) and Dunaliella tertiolecta) with distinctly different fatty acid profiles were grown in continuous culture and fed to prawn larvae (Penaeus japonicus, P. semisulcatus and P. monodon) as monospecific diets. The best two diets (C. muelleri and T. suecica) were also fed as a mixed diet. Experiments were run until the larvae fed the control diet of C. muelleri metamorphosed to Mysis 1. The survival and development (i.e. performance) of the larvae were affected by algal diet, and the diets were ranked in the order of decreasing nutritional value: C. muelleri ≥ T. suecica > T-iso > D. tertiolecta. Larvae fed a mixed diet of C. muelleri and T. suecica (2:3 by dry weight) performed as well or better than those fed C. muelleri, and the performance of both these groups of larvae was better than those fed T. suecica. The lipid and carbohydrate compositions of the algae had little or no effect on the lipid and carbohydrate compositions of the larvae or their performance. However, the larvae that performed best (i.e. those fed C. muelleri) had significantly more lipid and carbohydrate than those that performed worst (i.e. those fed D. tertiolecta). Larvae fed C. muelleri or the mixed-algae diet had higher proportions of the essential fatty acids eicosapentaenoic acid [EPA, 20:5(n-3)] and arachidonic acid [ARA, 20:4(n-6)] than the larvae fed on other diets. Furthermore, the larvae fed T. suecica, which showed intermediate performance between larvae fed C. muelleri and T-iso or D. tertiolecta, also had higher proportions of EPA and ARA. Both C. muelleri and T. suecica contained EPA and ARA, but T-iso and D. tertiolecta did not, except for trace amounts of EPA in T-iso. The fatty acid ARA appears to be much more important in the diet of larval prawns than has so far been considered. The level of the essential fatty acid docosahexaenoic acid [DHA, 22:6(n-3)] in the algal diet and the larvae was not related to the performance of the larvae; only C. muelleri and T-iso contained DHA. However, the nauplii contained large proportions of DHA, suggesting that these were sufficient to meet the larval requirements for DHA during their development to Mysis 1. Mixed-algae diets could improve the performance of larvae by providing a more comprehensive range of fatty acids. Received: 22 April 1998 / Accepted: 3 December 1998     

Symbiotic anemones can grow when starved: nitrogen budget for Anemonia viridis in ammonium-supplemented seawater

The ability of endosymbioses between anthozoans and dinoflagellate algae (zooxanthellae) to retain excretory nitrogen and take up ammonium from seawater has been well documented. However, the quantitative importance of these processes to the nitrogen budget of such symbioses is poorly understood. When starved symbiotic Anemonia viridis were incubated in a flow-through system in seawater supplemented with 20 μM ammonium for 91 d under a light regime of 12 h light at 150 μmol photons m⁻² s⁻¹ and 12 h darkness, they showed a mean net growth of 0.197% of their initial weight per day. Control anemones in unsupplemented seawater with an ammonium concentration of <1 μM lost weight by a mean of 0.263% of their initial weight per day. Attempts to construct a nitrogen budget showed that, over a 14 d period, ≃40% of the ammonium taken up could be accounted for by growth of zooxanthellae. It was assumed that the remainder was translocated from zooxanthellae to host. However, since the budget does not balance, only 60% of the growth of host tissue was accounted for by this translocation. The value for host excretory nitrogen which was recycled to the symbionts equalled that taken in by ammonium uptake from the supplemented seawater, indicating the importance of nitrogen retention to the symbiotic association. Received: 23 December 1997 / Accepted: 12 September 1998     

Preliminary report on the digestive enzymes present in the digestive gland ofPerna viridis

Quantitative data on digestive enzymes of bivalves are very limited and so this study was performed to obtain such information, and to find out whether some of these enzymes are well adapted to conditions in the digestive gland. The green musselPerna viridis was obtained from Clementi West Market in 1985. The results show that various carbohydrases are present in the digestive gland. The activities of-amylase, cellulase and-glucosidase are higher than that of other carbohydrases, whilst the activities of both lipase and protease, especially the former, are low. The pH optima of the following enzymes are also given:-amylase = 5.8; laminarinase = 5.5;-glucosidase = 5.8; trehalase = 4.8;-glucosidase = 5.5. The only enzyme which was not well adapted to conditions in the digestive gland (pH 6.2) was trehalase. The wide spectrum of enzymes present in this bivalve indicates that it has the ability to utilize a wide range of nutrients efficiently.     

Size-specific changes in amount of lipid and daily growth rate of early larval sardine, Sardinops melanostictus, in the main Kuroshio Current and its offshore waters off eastern Japan

The size-specific nutritional conditions of larval sardines, Sardinops melanostictus, from the main Kuroshio Current and its offshore waters off eastern Japan were assessed by lipid analysis. A rapid lipid analysis technique (diagnostic kits for human serum lipids) was used to measure the different lipid components of individual sardine larvae as indicators of their nutritional condition. Size-specific growth trajectories of individual larvae were estimated by the biological intercept method, and the recent daily growth rate of standard length (SL) was calculated from the 3 d outer increment width on the otolith. Relationships between the amount of larval phospholipid (PL; tissue weight indicator) and SL, and the recent daily growth rate of larva (Gr) and SL, could be expressed by the equations PL = 0.459 SL ^1.77 and Gr = 0.0809 SL − 0.341, respectively. There was no notable difference in these values between the two survey areas. The relationship between the amount of triglyceride (TG) and SL could be expressed allometrically (TG = 0.013 SL ^2.63). The relationship between the index of starvation tolerance (TG/PL) and SL could be expressed by the allometric equation TG/PL = 0.0288 SL ^0.865, suggesting that larger sardine have a higher starvation tolerance than smaller sardine. The TG of the 8 to 9 mm SL size-class larvae in the offshore area was higher than in the main Kuroshio Current. To test whether the TG for each larval sardine in the 8 to 9 mm SL size-class could be correlated with variables (temperature, chlorophyll a and distribution density of the sardine larvae) measured at the sampling stations, correlation analyses were performed. A highly significant negative correlation between TG and distribution density of the sardine larvae was found. A density-dependence effect seemed to influence the fluctuation of the larval storage energy component for short-term needs. Received: 12 March 1998 / Accepted: 26 March 1999     

Home range, activity and distribution patterns of a temperate rocky-reef fish, Cheilodactylus fuscus

Red morwong (Cheilodactylus fuscus; Cheilodactylidae: Perciformes), are large temperate rocky reef fish that are patchily distributed in local aggregations of 3 to >100 fish. The home ranges and aggregating behaviour of red morwong were investigated using external tags (n = 114, over 35 mo) and ultrasonic transmitters (n = 9 over 20 d) at coastal and estuarine sites in south-east Australia. Adult red morwong had a mean home range during the day of 1865 m² (SE = 268). Night movement, determined by ultrasonic telemetry, indicated a significantly greater mean home range of 3639 m² (SE = 416). Home range did not differ significantly with size or sex. Fish movements were greatest and aggregation sizes varied most during the non-reproductive mid-summer period. Tagged fish (N = 20) displaced 200 to 900 m returned in 1 to 3 d to the point of capture, often traversing open sand habitat and other aggregations. During the day, fish were commonly found on bouldered habitat. During the night, these site-specific aggregations fragmented as fish dispersed over a variety of substrata, with crepuscular peaks in activity. Therefore, estimates of habitat-specificity and feeding patterns collected only by day may give misleading results. Received: 28 June 1996 / Accepted: 8 July 1998     

Differences of two polychaete species reflected in enzyme activities

Polychaetes constitute most of the benthic macroinvertebrates in estuarine and coastal environments. We investigated the utilization of organic matter in two polychaete species, Capitella sp. I and Perinereis nuntia brevicirris, living in different coastal habitats. The protease activity of Capitella sp. I (89.7 μg mg⁻¹) was about 10 times that of P. nuntia brevicirris (8.0 μg mg⁻¹). High cellulase (endo-β-1,4-glucanase) activity was detected in P. nuntia brevicirris (3.2 μg mg⁻¹), whereas scarcely any was detected in Capitella sp. I. We isolated cDNA clones of protease mRNA from Capitella sp. I and of cellulase mRNA from P. nuntia brevicirris. The high protease activity of Capitella sp. I enabled it to survive in the sediment under a fish farm, where it degrades organic matter. In contrast, the high cellulase activity of the estuary-dwelling P. nuntia brevicirris allowed it to degrade organic matter originating from terrestrial areas.     

Impact of starvation and of feeding algal and artificial diets on the lipid content and composition of juvenile oysters (Crassostrea gigas) and clams (Tapes philippinarum)

The present study evaluated the effect of starvation and of feeding various algal and lipid-supplemented diets on the lipid content and lipid class distribution in the polar and neutral lipids of early juvenile oysters (Crassostrea gigas) and clams (Tapes philippinarum L.). T. philippinarum was starved, fed a mixed algal diet [Tetraselmis suecica and Isochrysis galbana (clone T-Iso)] or solely T. suecica at three different feeding rations, either supplemented or not supplemented with lipid emulsions. C. gigas was fed T. suecica with and without the supplementation of lipid emulsions or liposomes. When T. philippinarum and C. gigas were fed solely T. suecica, no qualitative and only minor quantitative differences were observed between the lipid class profile of both species. The major neutral lipids were triglycerides (TAG) and free sterols plus diglycerides (FS+DAG), whereas the polar lipids were dominated by phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol plus ceramideaminoethanol phosphonate (PI+CAEP). An increase of the algal feeding ration resulted in an increase of the lipid content of spat which was associated with a significant augmentation of the TAG level (as percentage of the total lipids and as percentage of the neutral lipids). Lipid supplementation evoked a similar though more pronounced effect. Starvation resulted in a significant decline of the lipid content and a complete depletion of the TAG reserve. Contrary to the neutral lipids (NL), the relative proportion (percentage of total polar lipids, PL) of the individual PL classes was hardly affected by the diet. The importance of lipid and TAG reserves in early juveniles is discussed. Received: 15 October 1999 / Accepted: 15 February 2000