Larval rearing of the giant Azorean barnacle,Megabalanus azoricus (Pilsbry, 1916): feeding trials,larval development and settlement on artificial substrata

A series of experiments were conducted to obtain an efficient larval rearing protocol for Megabalanus azoricus. The first part of this study investigates the effect of microalgae‐based diets on survival and larval development. Mono and mixed‐diets were tested at 20 ± 1°C, in a sequence of 11‐day feeding experiments. The second part presents a preliminary study on the influence of a biofilm on recruitment and use of oyster spat collectors in a mass rearing system. A photographic record of larval development and a brief reference to the diagnostic features that enable quick larval staging are also presented, along with morphometric measurements. Of the microalgae tested (Chaetoceros sp., Chloromonas sp., Dunaliella sp., T‐Isochrysis sp. and Skeletonema sp.) the mixed‐diet Skeletonema sp. with T‐Isocrysis sp. showed the highest survival percentages: total survival ranged from 79.7 to 85.7% and 69.7–80.0% of nauplii were in stage VI after 11 days of rearing. Cypris were also present, but only represented 5.3% of the survivors at most. In the mass rearing system juveniles were found settled in the collectors after 25 days, at 20 ± 1°C. However recruitment was less than 1%. Preliminary results showed no settlement preference towards collectors with biofilm. Nevertheless, this study provides the first record of M. azoricus settlement under laboratorial conditions and represent a starting point for future larval rearing studies.     

Growth, filtration and ingestion rate of the rotifer Brachionus plicatilis fed with large (Asteromonas gracilis) and small (Chlorella sp.) celled algal species

The rotifer Brachionus plicatilis was fed in experimental conditions with a small celled (2–5 μm) Chlorella sp. and a large celled (16–22 μm) Asteromonas gracilis algae. The specific growth rate (SGR) of rotifers fed Asteromonas (maximum 0.79) was statistically higher than that for rotifers fed Chlorella (maximium 0.61). The filtration and ingestion rates using different rotifer and algal densities exhibited certain maxima depending on the species, the cell density and the condition of the rotifers. The filtration rate was higher with Asteromonas and, although ingestion rate was lower than with Chlorella, the ingestion in terms of cell volume was 10‐fold higher. It seems that B. plicatilis ingests the larger cell diameter algal species more efficiently than the smaller species that is usually used for its mass culture.     

Ingestion and digestion index of catarina scallop Argopecten ventricosus-circularis, Sowerby II, 1842, veliger larvae with ten microalgae species

The initial ingestion age and size and the digestive index of Argopecten ventricosus-circularis Sowerby II, 1842 veliger larvae were investigated using epifluorescence microscopy. Ten microalgae were tested as feed during the first 7 days of life at this stage. Results of ingestion experiments showed there was a significant positive correlation between the smaller geometric dimension of the algae and the larval age and size of initial ingestion. Out of the 10 species offered, only Nannochloris oculata (NANO), Isochrysis galbana (TISO), Monochrysis lutheri (MONO), Chaetoceros muelleri (CM), Chaetoceros calcitrans (CC), Thalassiosira pseudonana clone 3H (3H), and Phaeodactylum tricornutum (PHT) were ingested. No ingestion was observed in Thalassiosira pseudonana clone s78 (S78), Tetraselmis suecica (TETRA) and Dunaliella tertiolecta (DUN) during the experiment. The digestion index of the larvae, defined as the degree of algal digestion at the third hour after ingestion, increased with larval age and size with TISO and MONO. With CC, 3H and CM, the digestion index was lower and no digestion was observed with NANO and PHT.     

Application of prospective probionts at early stages of Atlantic cod (Gadus morhua L.) rearing

This work aimed at validating the use of two prospective probionts (Arthrobacter sp. and Enterococcus sp.) at early stages of cod (Gadus morhua L.) rearing. Ova at late post‐fertilized stage and larvae during their first 4 weeks of life were bathed with both probionts, isolated previously from the cod‐rearing environment. This treatment was compared with groups fed rotifers supplemented with a commercial probiotic (Remus^®) and those untreated. Microbiological analyses (total viable counts, presumptive Vibrio and lactic acid bacteria) were performed in rearing systems and larval survival, growth and development were assessed. Larval development was evaluated by proteolytic activity of larval lysates and immunological analysis of important proteins: apolipoprotein A‐I, haemoglobin, C‐reactive protein, C3 and cod serum proteins. Bacterial bathing led to a significantly higher larval weight, length and culturable microbial load in larval gastrointestinal (GI) tract when compared with the control and Remus groups. Development occurred earlier in bathed larvae. However, their survival was negatively affected compared with the control group, but was significantly higher than for the Remus group. The non‐pathogenicity of both probionts was demonstrated by intraperitoneal injection of 13 g cod juveniles. The results suggest that Arthrobacter and Enterococcus probionts affected the larval GI microbiota and contributed to growth, development and digestion, either directly or indirectly.     

Meiotic chromosome configurations in triploid and heteroploid mosaic males of Crassostrea gigas and Crassostrea ariakensis

Reversion from triploids to diploids or heteroploid mosaics may make the revertants recover reproductive ability and lose their aquacultural advantages. Meiotic chromosomes in triploids and mosaics of Crassostrea gigas and Crassostrea ariakensis were studied. Pachytene spermatocytes showed incomplete trivalent formation, varying 64–72% in C. gigas and 62–69% in C. ariakensis. At metaphase I, trivalents, bivalents and univalents occurred in various combinations. Trivalents occurred in all spermatocytes, ranged 6–10 per spermatocyte and averaged 8.53–8.97 in C. gigas and 8.49–8.95 in C. ariakensis. Univalents and bivalents appeared in 59.2–72.1% of the spermatocytes in C. gigas, and ranged 0.77–1.39 and 1.01–1.51 per spermatocyte respectively. In C. ariakensis, they occurred in 57.5–81% of the spermatocytes, and ranged 0.9–1.32 and 1.03–1.35 respectively. The most common trivalent was formed by a bivalent with the third chromosome attaching to its side to form ‘long‐tail‐cross’ or ‘t’ or ‘frying‐pan’‐shaped configurations that constituted 65.7% and 59.9% of the trivalents in C. gigas and C. ariakensis respectively. Other kinds of trivalent associations included tandem chains (14.7%, 16.5%), closed circles or triangles (5.3%, 9.8%), convergent ‘T’ or ‘Y’ (5.8%, 7.4%) and unclassified configurations (8.5%, 6.4%). Diploid spermatocytes were not observed from mosaics, although they showed considerable proportions of ‘diploid’ cells in their gills.     

Influence of one selected Tisochrysis lutea strain rich in lipids on Crassostrea gigas larval development and biochemical composition

Effects of a remarkably high overall lipid Tisochrysis lutea strain (T+) upon gross biochemical composition, fatty acid (FA), sterol and lipid class composition of Crassostrea gigas larvae were evaluated and compared with a normal strain of Tisochrysis lutea (T) and the diatom Chaetoceros neogracile (Cg). In a first experiment, the influence of different single diets (T, T+ and Cg) and a bispecific diet (TCg) was studied, whereas, effects of monospecific diets (T and T+) and bispecific diets (TCg and T+Cg) were evaluated in a second experiment. The strain T+ was very rich in triglycerides (TAG: 93–95% of total neutral lipids), saturated FA (45%), monounsaturated FA (31–33%) and total fatty acids (4.0–4.7 pg cell^?1). Larval oyster survival and growth rate were positively correlated with 18:1n‐7 and 20:1n‐7, in storage lipids (SL), and negatively related to 14:0, 18:1n‐9, 20:1n‐9, 20:4n‐6 and trans‐22‐dehydrocholesterol in membrane lipids (ML). Surprisingly, only the essential fatty acid 20:5n‐3 in SL was correlated positively with larval survival. Correlations suggest that physiological disruption by overabundance of TAG, FFA and certain fatty acids in larvae fed T+ was largely responsible for the poor performance of these larvae. ‘High‐lipid’ strains of microalgae, without regard to qualitative lipid composition, do not always improve bivalve larval performance.     

Optimizing initial feeding of the Pike silverside Chirostoma estor: oil droplet depletion,point of no return,growth and fatty acid utilization in larvae fed enriched rotifers

The point of no return (PNR) and disappearance of the oil droplet were measured in Chirostoma estor larvae as a function of the time of first feeding. In a separate trial, growth and survival of larvae fed rotifers enriched with Chlorella sp., cod liver oil and corn oil were assessed. Fatty acid and lipid composition of eggs, oil droplets, egg yolk, feed and larvae were also evaluated. The PNR was found between 7 and 8 days posthatching (dph). Total oil droplet depletion occurred between 7 and 11 dph, depending on the time of first feeding. Best growth and survival were obtained in larvae fed with Chlorella‐enriched rotifers, followed by those fed cod liver oil‐enriched rotifers. In larvae fed corn oil, Chlorella and cod liver oil‐enriched rotifers, total oil droplet depletion took place on days 9, 10 and 11, respectively. There was a direct relationship between presence and duration of oil droplets and the survival of larvae under different starvation conditions. The feed source could prolong the existence of the oil droplet depending on particular dietary supply of essential fatty acids; the time of its disappearance could be a useful indicator of larval vigour and health status.     

The effect of temperature on physiological energetics of a fast‐growing selective strain and a hatchery population of the Pacific oyster (Crassostrea gigas)

Growth is an important aquaculture trait, which has been one of the most important traits for genetic improvement in aquatic species. Numerous fast‐growing domesticated aquaculture organisms are being cultured, but the physiological processes underlying growth remain largely unexplored. The Pacific oyster (Crassostrea gigas) is one of the most widely farmed bivalves. In order to improve the productivity traits, a fast‐growing selective strain of C. gigas was established. In this study, we compared physiological energetics of the fast‐growing selective strain and an unselected hatchery population of C. gigas under different conditions of temperature. Components of energy gain (clearance rates, ingestion rates, absorption efficiency and absorption rates), energy loss (metabolic rates) and resulting energy budget (scope for growth) were analysed at four temperatures (15, 20, 25 and 30°C). The results indicated that selective breeding oysters had a higher energy gain compared to unselected oysters, but the selected strain and hatchery population of oysters did not significantly differ in metabolic rates at all tested temperatures (p > 0.05). The enhanced energy gain resulted in almost two times higher scope for growth in selective breeding oysters compared with unselected oysters. Physiological characteristics of C. gigas were greatly affected by temperature and showed a fluctuation with the increase in temperatures. The selective breeding oysters outperformed those of hatchery population in growth performance by an increased energy gain along with a similar energy loss. This study provided insight into physiological energetics accounting for growth improvements in fast‐growing selective strain of C. gigas.     

Effect of using Spirulina and Chlorella as feed additives for elevating immunity status of Nile tilapia experimentally infected with Aeromonas hydrophila

The present trial was performed to evaluate the effect of dietary incorporation of dried Spirulina, Chlorella and their mixture on the immune status of Nile tilapia (Oreochromis niloticus) before and after artificial infection with pathogenic bacterium Aeromonas hydrophila (A. hydrophila). Two hundred and forty fish were divided into four groups: (a) a control group fed on a basal diet only, (b) a second group fed on a diet containing Spirulina 15%, (c) a third group fed on a diet containing Chlorella 15%, and (d) a fourth group fed on a diet containing a mixture of both Spirulina 15% and Chlorella 15%. At the end of the experiment (9 weeks), the four groups were experimentally infected with A. hydrophila for 7 days. Antioxidant enzymes, lysozyme and bactericidal activities and histopathological changes were determined just before the challenge test and 7 days post‐challenge. Significant (p ≤ 0.05) increases in fish body protein% before the challenge test and increases in serum antioxidant enzymes, lysozyme and bactericidal activity in the Chlorella and algal mixture groups before and after the challenge test were observed. Spirulina, Chlorella and their mixture groups significantly decreased serum malondialdehyde compared to the control group before and after the challenge test. Using Spirulina, Chlorella and their mixture mitigated the necrotic and degenerative changes induced by A. hydrophila and revealed well‐developed and multiple melanomacrophage centres. Thus, dietary Spirulina, Chlorella and their mixture inclusion in Nile tilapia fish proved to have a protective effect against A. hydrophila infection.     

Growth and Survival of Larval Sandfish,Holothuria scabra (Echinodermata: Holothuroidea), Fed Different Microalgae

This study assessed the nutritional value of four species of microalgae for larval sandfish, Holothuria scabra (Echinodermata: Holothuroidea). Two‐day‐old larvae were fed either a single diatom (Chaetoceros muelleri, Chaetoceros calcitrans), flagellate (Isochrysis aff. galbana (T‐ISO), Pavlova salina), or a ternary microalgal diet (TMD) comprised of 40% C. muelleri, 40% T‐ISO, and 20% P. salina for 12 d. On Days 4, 6, and 8 length of larvae fed either C. muelleri, C. calcitrans, or the TMD was significantly greater than in T‐ISO and P. salina fed larvae as well as the unfed control larvae. T‐ISO fed larvae had a significantly greater length compared to unfed larvae on Days 4, 6, and 8; however, larvae fed P. salina had a significantly greater length compared to unfed larvae on Days 6 and 8 only. Survival of larvae fed either C. muelleri or the TMD was significantly better than the flagellate fed and unfed larvae on Days 4, 8, and 12. There was no significant difference between survival of unfed larvae and larvae fed either T‐ISO or P. salina during the 12‐d larval culture period. On Day 12, larvae fed either a diatom or the TMD developed in excess of 84% competent doliolariae. Larvae fed C. muelleri had a significantly higher percentage competent doliolariae than larvae from all other treatments except the TMD. Neither larvae fed either T‐ISO or P. salina nor unfed larvae developed any competent doliolariae. In conclusion, this study showed C. muelleri to be the most effective of the four microalgae tested as a single microalgal diet for larval H. scabra and, as such, C. muelleri will be used as a reference diet in future studies.     

Abnormal morphology in megalopae of the swimming crab,Portunus trituberculatus during seed production: causes and prevention

The occurrence of morphologically immature megalopae, which retain zoeal features such as dorsal spines and furcae of telson, is closely correlated with larval mass mortality during seed production of the swimming crab Portunus trituberculatus in Japanese hatcheries. To determine the cause of immature megalopal morphology, zoeae were reared with various supplementary schedules and density of diets (rotifer, Artemia and phytoplanktons including Chlorella vulgaris and Nannochloropsis oculata). In addition, to assess the relationship between immature morphology and endocrine control, the effect of causative dietary factor was compared with the effect of eyestalk ablation at various timing during zoeal development. Megalopal immature morphology was more distinct in Chlorella‐supplemented groups than in Nannochloropsis‐supplemented groups. High density Chlorella supplementation was associated with the highest incidence of immaturity and resulted in larval mass mortality. The premoult of the third zoeal stage was identified as a critical period at which Chlorella supplementation led to the highest incidence of immaturity. This critical period coincided with the critical period at which larval metamorphosis was regulated by the eyestalk neurosecretory system. Our results suggested that the occurrence of immature megalopal morphology under culture conditions is most likely caused by phytoplankton (especially, Chlorella) supplementation, which disrupts the endocrine regulation. On the basis of our results, we successfully prevented the occurrence of immature megalopal morphology in 500 L tanks by excluding the influence of phytoplankton before the critical period (i.e. discontinuing phytoplankton supplementation and supplying rotifer cultured with non‐phytoplankton materials).     

Effects of on‐demand feeding on sea urchin larvae (Paracentrotus lividus; Lamarck, 1816), development,survival and microalgae utilization

This study compared the growth of sea urchin Paracentrotus lividus larvae cultured using two different rearing methods: a variable method based on a variable amount of feed (microalgae) and seawater exchange (30% or 50%) established according to the phytoplankton concentration in the larval cultures and a fixed method characterized by a fixed amount of feed and seawater exchange. Three microalgae diets, Isochrysis sp. (Tahitian strain, T‐Iso), Chaetoceros gracilis and a 50:50 mixed diet, were tested with both rearing methods. Larval development and survival were assessed at the 6‐arm pluteus stage (P6), competence (Cp) and metamorphosis (Mt). Data showed that the variable method reduced the requirements for phytoplankton and seawater exchange. Indeed, through the optimization of feed rations, it was possible to reduce the production of debris and settled phytoplankton, minimizing the need for water exchanges. Higher larval survival resulted at Cp and Mt stages for those reared with the variable method as opposed to the fixed one. Survival and development were also influenced by the tested dietary treatments: at Mt stage, the mixed diet resulted in a higher larval survival (63.3 ± 8.9%) than T‐Iso (19.7 ± 12.1%) and C. gracilis (23.4 ± 15.1%) (P < 0.05). These results suggest that the use of the variable method improves the larval survival and development and also it reduces resource consumption (phytoplankton, seawater use and work effort), which in turn could potentially improve the hatchery production of P. lividus.     

Selectivity of the rotifer Brachionus plicatilis fed mixtures of algal species with various cell volumes and cell densities

The ability for food selectivity of rotifer Brachionus plicatiliswas studied in the laboratory by feeding mixtures of microalgae with various cell volumes. Chlorella sp. (≈22 µm³) was the reference algal species, and Asteromonas gracilis, (Chlorophyta) (≈2150 µm³), Tetraselmis suesica (≈268 µm³), Dunaliella salina (≈52 µm³) and Chaetoceros sp. (≈150 µm³) the experimental species. Each was mixed with Chlorella and fed in three experiments. In the first experiment, filtration and ingestion rates of rotifers each fed with algae revealed that the highest values were measured with the mixture of Chlorella + Asteromonas, and the lowest for Chlorella + Chaetoceros. In the second and the third experiments, by using several combinations of algal densities with the mixture of Chlorella + Asteromonas, a selectivity ability of the rotifers for Asteromonas was found. A hypothesis is presented that accounts for the preference of rotifers for Asteromonas, which is suggested as a new candidate species for use in live food production of fish hatcheries.     

Food value of Brachionus plicatilis fed three selected algal species as live food for milkfish, Chanos chanos Forsskal, fry production

Abstract. The effects of three selected algal species, Tetraselmis tetrahele, Isochrysis galbana and marine Chlorella sp. on the population growth of Brachionus plicatilis was evaluated after 3, 5 and 7 days of culture. The rotifers fed on T. tetrahele showed superior growth with mean peak density of 92.5 individuals per ml to those fed on I. galbana (48.2 individuals per ml) and Chlorella sp. (47.2 individuals per ml) in 5 days. In another experiment, milkfish. Chanos chanos Forsskal, fry with initial mean body weight and standard length of 6.26 ± 1.07mg and 13.04 ± 1.40mm, respectively, were fed B. plicatilis reared on the three algal species for 30 days. Results showed that growth, measured in terms of gain in weight and length, was significantly different (P > 0.05) among treatments. Fastest growth was obtained in fry fed B. plicatilis cultured on T. tetrahele with mean gains in weight and length of 61.48 ± 8.40mg and 9.37 ± 1.68 mm, respectively, followed by fry fed B. plicatilis reared on I. galbana (43.56 ± 8.35mg and 6.25 ± 0.48mm). Milkfish fry fed B. plicatilis cultured on Chlorella sp. gave the poorest growth with mean gains in weight and length of 11.05 ± 1.82mg and 2.33 ± 0.29mm, respectively. Percentage survival ranged from 93.1 to 95.3 percent and was not significantly different between treatments (P < 0.05). The differences in population growth of B. plicatilis and consequently growth of milkfish fry was almost certainly attributed to the differences observed in the biological values of their algal diet, and in particular their protein and lipid contents. T. tetrahele and I. galbana improved the food value of B. plicatilis as live food for the milkfish fry. Their high dietary value was related to the higher algal protein and fat levels and contents of n-3 HUFA particularly 20:5n-3 and 22:6n-3 compared to B. plicatilis cultured on Chlorella sp. The results of the present study point to T. tetrahele as the most suitable food for the mass culture of the rotifer, B. plicatilis.     

Use of marine yeasts as an available diet for mass cultures of Moina macrocopa

A 4‐week feeding trial was conducted to test the use of marine yeasts purified from seawater and sediments as a dietary source in cultivating a cladoceran, Moina macrocopa, a potential alternative live food for fish larvae. Optimal growth conditions of two yeast strains were obtained for NaCl concentration, pH and temperature. Moina macrocopa was cultivated using different diets: marine yeasts (Debaryomyces hansenii Yeast‐14 and Candida austromarina Yeast‐16) and a commercial diet (Erythrobacter sp. Sπ‐I). The essential amino acids of both the yeast strains were nearly as great as those in M. microcopa. Further, the yeast‐fed M. macrocopa had essential amino acid profiles similar to the documented values for rotifers and Artemia enriched in microalgae and commercial diets. Erythrobacter sp. Sπ‐I lacked n‐3 polyunsaturated fatty acids, 20:5n‐3 and 22:6n‐3, which were also low but detected in both yeasts. An increase in the 20:5n‐3 [eicosapentaenoic acid (EPA)] and 22:6n‐3 [docosahexaenoic acid (DHA)] levels, compared with the levels in yeast strains themselves, was more pronounced in the 22:6n‐3 level of Moina fed the C. austromarina, resulting in a high DHA:EPA ratio. When the Moina diets were switched, their δ¹³C values shifted gradually towards the values of the switched diets. Diet switch from Erythrobacter sp. Sπ‐I to C. austromarina Yeast‐16 resulted in a more rapid turnover of Moina tissue carbon than that in the inverse case. When fed a mixed diet, the δ¹³C values of Moina tissue approached the value of marine yeasts immediately. These temporal changes in the δ¹³C values of Moina tissue indicate the preferential ingestion of marine yeasts and a selective assimilation of the carbon originated from marine yeasts. These findings suggest that marine yeasts, particularly C. austromarina Yeast‐16, are highly available to mass cultures of M. macrocopa, providing better nutritional and dietary values than the commercial diet (Erythrobacter sp. Sπ‐I).     

The effects of dietary supplements of polyunsaturated fatty acid on pearl oyster,Pinctada margaritifera L., gonad composition and reproductive output

Black‐lip pearl oyster, Pinctada margaritifera broodstock was collected from the wild. Egg production, hatching rate and larval development were compared between oysters induced to spawn within 2 days after collection in the wild (T1), oysters fed a pure microalgae diet during 24 days before spawning (T2) and oysters fed the same microalgal diet in which 10% of the algae were replaced with 2 μm polyunsaturated fatty acid (PUFA)‐rich microspheres (T3). Administration of lipid microspheres resulted in larger sized eggs, a higher percentage of D‐larvae and larger sized 48‐h‐old larvae (P<0.05). The total and neutral lipid contents of the gonad increased after oysters were fed with microalgae only or with supplementary diet. The major neutral and polar fractions of saturated fatty acid (SFA) were 16C and 18C fatty acids, and not influenced by the diet (P>0.05). The gonads of oysters fed supplementary PUFA contained more docosahexaenoic acid (DHA) and less monounsaturated fatty acids. Higher level of DHA in gonads of T3 was associated with oogenesis and embryogenesis success. The n‐3/n‐6 ratio in the neutral lipid fraction provides a good indication of the spawning condition and predicting egg size and hatching rate.     

Ingestion,fecundity and population growth of Harpacticus sp. (Harpacticoida,copepod) fed on five species of algae

We investigated the ingestion, fecundity and population growth of Harpacticus sp. fed on monodiet or mixed diets to evaluate the effects of different algae on Harpacticus sp. Harpacticus sp. fed on diatoms (Skeletonema costatum and Chaetoceros curvisetus) had higher ingestion and pellet production. Time to attain 100% in proportions of gravid females differed, with quickest to slowest: S. costatum, C. curvisetus, Gymnodinium sp. and Heterosigma akashiwo, with the exception of Prymnesium parvum (≤16.67%). S. costatum or C. curvisetus produced higher populations than the other three diets, supported complete development to adulthood, and resulted in doubling copepod population within four days, while no population growth occurred for the other three diets. Mixed‐diet experiments showed that egg production and gross growth efficiencies reduced significantly when fed on H. akashiwo, Gymnodinium sp. or P. parvum mixed with Isochrysis galbana, in comparison with 100% I. galbana. Thus, S. costatum and C. curvisetus were beneficial foods while the other three diets were potentially toxic for Harpacticus sp. The data in this article provide further recognition of nutrient deficiency or toxicity of different algae on copepods.     

Ingestion and digestion of 10 species of microalgae by winged pearl oyster Pteria sterna (Gould, 1851) larvae

Ten species of microalgae were tested for ingestion and digestion in Pteria sterna larvae using epifluorescence microscopy to choose an appropriate diet. An experiment was conducted using 2, 4, 5 (straight-hinge) 10 and 22 (umbo stage) day old larvae. Larvae were stocked in 150 ml flasks at 30 ml⁻¹ and fed 100,000 algal cells ml⁻¹ of each species individually. Larvae were fed for 1 h and then were observed under the microscope to detect ingestion; larvae were then sieved and placed in flasks containing filtered seawater and were observed after 1 and 2 h to analyse digestion for the microalgae ingested. Out of the 10 species administered, only Nannochloris sp., Pavlova lutheri and Isochrysis aff. galbana (T-ISO) were ingested, and only the last two species were digested. No ingestion of Phaeodactylum tricornutum, Chaetoceros muelleri, Ch. calcitrans, Thalassiosira weisflogii, Dunaliella salina, Tetraselmis tetrathele and T. suecica was evident at any stage of larval development tested. Only T-ISO and P. lutheri should be used for larval rearing of P. sterna until other species are identified as suitable (ingested and digested) for this species.     

Insights on the association between somatic aneuploidy and ostreid herpesvirus 1 detection in the oysters Crassostrea gigas,C. angulata and their F1 hybrids

Cytogenetic abnormalities associated with viral infections, including from viruses of the Herpesvirales order, have been reported in vertebrate species. Ostreid herpesvirus 1 (OsHV‐1) has been detected worldwide during mortality outbreaks of the Pacific oyster Crassostrea gigas. On the other hand, a high proportion of aneuploid cells in somatic tissues have been observed in C. gigas. In this study, we analysed the putative association between aneuploidy levels and the detection of OsHV‐1 in gills of C. gigas, the Portuguese oyster C. angulata and their F1 hybrids cultured in Ria Formosa (Portugal). OsHV‐1 was detected by PCR in 5.4% of the total of oysters analysed (n = 111) namely in 11.1%, 8.0% and 1.7% of C. gigas, C. angulata and F1 hybrid respectively. Sequencing analysis of a viral fragment amplified with the C2/C6 primer pair revealed a high similarity with the OsHV‐1 reference type. Moreover, in situ hybridization confirmed the presence of OsHV‐1 in gill tissue. Oysters where OsHV‐1 was detected had a significantly higher mean percentage of aneuploid cells (25%) than the ones where the virus was not detected (18%). However, the overall low percentage of positive samples contrasted with the high mean percentage of aneuploidy observed, with 50% of the oysters analysed showing a percentage of aneuploid cells between 20% and 30%. We hypothesize that somatic aneuploidy may adversely affect oysters making them more prone to OsHV‐1 infection, but the virus is unlikely to be the cause of somatic aneuploidy.     

Digestion of lipids and carbohydrates from microalgae (Chaetoceros muelleri Lemmermann and Isochrysis aff. galbana clone T‐ISO) in juvenile scallops (Pecten maximus L.)

The digestion of laminaran and neutral lipids from the microalgae Chaetoceros muelleri or mixtures of C. muelleri and Isochrysis galbana clone T‐ISO in 4‐mm scallops Pecten maximus were studied. The microalgae were cultivated at fixed low and high growth rate (5% and 60% of μ_max). The cultivation method clearly affected the lipid and carbohydrate contents, as well as their enzyme activity. Chaetoceros muelleri and T‐ISO grown at low growth rate exhibited a considerably higher content of lipids and carbohydrates than those algae grown at high growth rate. The specific activity of laminaranase and neutral lipase was the highest in C. muelleri grown at the low growth rate. Growth rate of scallops was best when they were fed a mono‐species diet of C. muelleri grown at low growth rate. The growth rate of the scallops was positively correlated to the dietary carbohydrate content as well as to the specific activity of neutral lipase and laminaranase in the diet. The capacity to digest carbohydrates (laminaran) might be limiting for the young scallops, and the content of carbohydrates and enzymatic activities of laminaran in the diet should be considered when formulating an optimal diet, both when the diet consist of live microalgae and formulated diets.