Improved techniques for rearing mud crab Scylla paramamosain (Estampador 1949) larvae

A series of rearing trials in small 1 L cones and large tanks of 30–100 L were carried out to develop optimal rearing techniques for mud crab (Scylla paramamosain) larvae. Using water exchange (discontinuous partial water renewal or continuous treatment through biofiltration) and micro‐algae (Chlorella or Chaetoceros) supplementation (daily supplementation at 0.1–0.2 million cells mL⁻¹ or maintenance at 1–2 millions cells mL⁻¹), six different types of rearing systems were tried. The combination of a green‐water batch system for early stages and a recirculating system with micro‐algae supplementation for later stages resulted in the best overall performance of the crab larvae. No clear effects of crab stocking density (50–200 larvae L⁻¹) and rotifer (30–60 rotifers mL⁻¹) and Artemia density (10–20 L⁻¹) were observed. A stocking density of 100–150 zoea 1 (Z1) L⁻¹, combined with rotifer of 30–45 mL⁻¹ for early stages and Artemia feeding at 10–15 nauplii mL⁻¹ for Z3–Z5 seemed to produce the best performance of S. paramamosain larvae. Optimal rations for crab larvae should, however, be adjusted depending on the species, larval stage, larval status, prey size, rearing system and techniques. A practical feeding schedule could be to increase live food density from 30 to 45 rotifers mL⁻¹ from Z1 to Z2 and increase the number of Artemia nauplii mL⁻¹ from 10 to 15 from Z3 to Z5. Bacterial disease remains one of the key factors underlying the high mortality in the zoea stages. Further research to develop safe prophylactic treatments is therefore warranted. Combined with proper live food enrichment techniques, application of these findings has sustained a survival rate from Z1 to crab 1–2 stages in large rearing tanks of 10–15% (maximum 30%).     

Effects of selenium supplementation on the antioxidation response in the rotifer (Brachionus plicatilis)

In the present study, the effects of selenium (Se) enrichment on the rotifer, Brachionus plicatilis, were investigated. Based on the results of six Se treatments obtained from triplicate experiments, we found that the efficiency of bioaccumulation by Se from non‐acid‐soluble sources was 62‐fold greater than that from acid‐soluble sources (Se‐Chlorella 15.6 ± 1.81% vs. selenite 0.25 ± 0.03%). However, supplementation of both acid‐soluble Se at low concentrations (≤40 µg per thousand rotifers) and Se‐Chlorella enhanced rotifer lifespan, increased the rate of spawning, inhibited lipid peroxidation, decreased levels of intracellular oxidization products and maintained the stability of mitochondrial membranes. These positive effects were correlated with the activities of antioxidant enzymes (e.g. catalase‐ and guaiacol‐dependent peroxidase) which increased along with the low Se concentrations (<80 µg per thousand rotifers). In contrast, the opposite effects were found when the rotifers were exposed to high Se concentrations (≥80 µg per thousand rotifers). The antioxidation and toxicity effects appeared to be correlated with the levels of total Se bioaccumulation. Above all, selenite, which is more toxic than Se‐chlorella, is less readily accumulated and biologically active. Se‐enriched rotifers fed by Se‐Chlorella are a fish larvae food supplement in aquaculture.     

Effects of docosahexaenoic acid on growth,survival and swim bladder inflation of larval amberjack ( Seriola dumerili, Risso)

The effect of docosahexaenoic acid (DHA) on the growth performance, survival and swim bladder inflation of larval Seriola dumerili during the rotifer feeding period was investigated in two feeding experiments. Amberjack larvae at 3 day post hatching were fed rotifers enriched with (1) freshwater C hlorella (Chlo), (2) a mixture (2:1, v/v) of Chlo and DHA‐enriched C hlorella (DHA‐Chlo), (3) DHA‐Chlo and (4) DHA‐Chlo and commercial DHA emulsion, in triplicate for 7 days. The average DHA contents of the rotifers were 0.0, 0.4, 1.0 and 1.9 mg g^?1 DM respectively. The survival rate was improved by the enrichment of rotifers with DHA‐Chlo alone, and DHA‐Chlo and emulsion. Growth and swim bladder inflation of fish fed rotifers enriched with DHA‐Chlo were significantly (P < 0.05) improved, however, with increased levels of DHA further improvement was not found. DHA content in the larval whole body proportionally increased with the DHA level in the rotifers. These results suggest that DHA enrichment of rotifers is effective to improve the growth, survival rate and swim bladder inflation of amberjack larvae. The DHA requirement of amberjack larvae is estimated to be 1.5 mg g^?1 on a dry matter basis of rotifers.     

Rotifers enriched with iodine,copper and manganese had no effect on larval cod (Gadus morhua) growth,mineral status or redox system gene mRNA levels

It currently remains unclear if rotifers contain sufficient mineral levels to meet larval fish requirements. In this study, rotifers were enriched with a commercial enrichment (control), or with additional iodine, iodine and copper, or iodine, copper and manganese, and the effects of feeding these rotifers to Atlantic cod (Gadus morhua) larvae from 3 to 18 days post hatch were investigated. Rotifer enrichment with minerals was successful, but Mn enrichment also increased rotifer zinc levels. No differences were observed between treatments in larval growth or survival, or in the mRNA levels in the majority of the redox system genes analysed. Only Zn levels increased in cod larvae in response to mineral enrichment of rotifers. Apart from Zn, little evidence was found to suggest that cod larvae require increased concentrations above the control rotifer levels of the essential elements studied here.     

Incidence of systemic granulomatosis is modulated by the feeding sequence and type of enrichment in meagre (Argyrosomus regius) larvae

Systemic granulomatosis is the most frequent disease in juvenile and adult meagre, but studies regarding the first appearance of granulomas in larvae do not exist. In order to evaluate this, meagre larvae were fed four different feeding regimes as follows: RS and RO (rotifer enriched with Easy DHA Selco or Ori‐Green from 3 to 30 dph respectively), RAS and RAO (rotifer enriched with Easy DHA Selco or Ori‐Green from 3 to 21 dph and Artemia enriched with Easy DHA Selco or Ori‐Green from 12 to 30 dph respectively). All treatments were also fed with commercial microdiet from 20 to 30 dph. At 30 dph weight, length, specific growth rate and survival were significantly higher in Artemia‐fed larvae, regardless of the enrichment. Microscopic first appearance of granulomas was observed in 20 dph larvae fed RS and RO. At 30 dph granulomas and thiobarbituric acid reactive substances (TBARS), values were significantly higher in RS and RO‐fed larvae than in RAS and RAO‐fed larvae. The results showed that granulomas first appeared in meagre larvae at 20 dph when fed rotifers only. Conversely, a reduced appearance of granulomas and lipid peroxidation occurs when Artemia is included in the feeding sequence reinforcing the hypothesis of a nutritional origin of the systemic granulomatosis.     

The effect of poly‐β‐hydroxybutyrate on the performance of Chinese mitten crab (Eriocheir sinensis Milne‐Edwards) zoea larvae

Two experiments were conducted regarding the effect of poly‐β‐hydroxybutyrate (PHB) on the survival and development of Chinese mitten crab (Eriocheir sinensis Milne‐Edwards) larvae. Different PHB delivery approaches (particulate, enriched in filter feeding prey, or a combination of both) and feeding levels were applied to crab larvae from the zoea 2 (Z2) up to the megalopa stage. Bio‐encapsulation into rotifers and Artemia at a PHB dose of 100 mg L^?1 seemed an efficient approach to deliver PHB to the larvae and resulted in a significantly enhanced survival, development rate and osmotic tolerance. The results indicate that PHB should only be applied as a feed additive rather than as a food source, and that the availability of an adequate amount of nutrients seems to be of major importance for the PHB to induce these beneficial effects.     

Effect of different algae used for enrichment of rotifers on growth, survival, and swim bladder inflation of larval amberjack Seriola dumerili

The effect of algae with different DHA contents used for the enrichment of rotifers on the growth performance, survival, and swim bladder inflation of larval amberjack Seriola dumerili was investigated. Rotifers were enriched with freshwater Chlorella vulgaris containing three levels of DHA (rotifer containing DHA 0.04, 0.60, 1.32?g DHA 100?g^?1 DM) and Nannochloropsis (0.04?g DHA 100?g^?1 DM; 2.54?g EPA 100?g^?1DM). The larvae were fed the enriched rotifers in triplicate from 3?days post-hatch for 7?days in static condition. The same algae used for rotifer enrichment were supplied to the larval tanks. Growth and survival rate of fish fed the rotifers enriched with Nannochloropsis were higher than those of fish fed the rotifers enriched with all three Chlorella treatments. Swim bladder inflation was lowest in fish fed the rotifers enriched with Nannochloropsis. The proportion of EPA was higher in fish fed the rotifers enriched with Nannochloropsis, while that of DHA increased proportionally with the DHA levels in the rotifers enriched with Chlorella. These results suggest that rotifers enriched with Nannochloropsis (EPA-rich rotifers) are effective to enhance growth and survival, but DHA instead of EPA is essential to improve the swim bladder inflation in amberjack larvae.     

Effect of feeding scheme and prey density on survival and development of Chinese mitten crab Eriocheir sinensis zoea larvae

The effect of feeding scheme and prey density on survival and development of Eriocheir sinensis zoea larvae was studied in three experiments. Different combinations and densities of rotifers (Brachionus rotundiformis) and newly hatched Artemia nauplii were fed to zoea larvae. Average survival at each stage, larval development (larval stage index, LSI), duration of zoeal stage and individual megalopa dry weight were compared among treatments. This study revealed that, under the experimental conditions, rotifers should be replaced with Artemia between the zoea 3 (Z3) and the zoea 4 (Z4) stage. The optimal rotifer feeding densities for zoea 1 (Z1) and zoea 2 (Z2) were 15 and 20 mL^?1 respectively, while the optimal Artemia feeding density for Z3, Z4 and zoea 5 (Z5) was 3, 5 and 8 mL^?1 respectively. Further trials in production scale are recommended.     

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.     

Replacement of fresh algae with commercial formulas to enrich rotifers in larval rearing of yellowtail kingfish Seriola lalandi (Valenciennes, 1833)

This study compared the efficacy of four products that are commonly used in hatchery for nutritional enhancement of rotifer Brachionus plicatilis as the starter food for yellowtail kingfish Seriola lalandi larvae. This experiment consisted of one fresh algae and three enrichment products: (1) Fresh algae were a mixture of Nannochloropsis and Isochrysis at 2:1 on a cell concentration basis; (2) S.presso, (Selco S.presso ^®, INVE Aquaculture); (3) Algamac 3050^® (Aquafauna, USA); (4) Nutrokol ^® (Nutra‐Kol, Australia). Survival rates of the fish fed rotifers enriched with fresh microalgae (40.69%) and S.presso (31.21%) were higher than those fed Algamac 3050 (10.31%). On 3 day post hatch (DPH), fish feeding incidence in the fresh algae treatment was significantly higher than that in other treatments. On 6 DPH, fish showed the lowest feeding incidence in the Algamac 3050 treatment. The methods of enrichment did not affect total lipid levels in either rotifer or fish larvae, but Algamac 3050 enrichment achieved the highest DHA/EPA ratio and lowest EPA/ARA ratio in both rotifers and fish larvae. This study indicates that fresh algae can be replaced by S.presso, but Algamac 3050 is not as good as other formula for rotifer enrichment in rearing yellowtail kingfish larvae in this system.     

Examination of a practical method for zinc enrichment of euryhaline rotifers (Brachionus plicatilis)

Larval growth and survival of marine finfish in mass seed production are affected by the nutritional value of live feeds such as rotifers and Artemia. Thus far, many studies have been conducted to develop effective methods for the enrichment of live feeds with essential fatty acids and vitamins. In this study, a practical method for enrichment of rotifers with zinc was investigated. Changes in the concentrations of other minerals when zinc was added to the rotifer-enrichment tanks were also studied. The mineral composition of rotifers and Chlorella after zinc enrichment revealed that the direct addition of zinc to the culture media was not effective because rotifers cannot efficiently accumulate waterborne zinc. The ability of Chlorella to absorb waterborne zinc is much higher than that of rotifers, and hence, zinc was pre-accumulated in Chlorella, which was then fed to the rotifers. The maximum zinc content of the rotifers was 585.0 μg g^? 1 (dry matter) when the rotifers were enriched with zinc alone. This zinc concentration is comparable to that found in natural zooplankton. In rotifers simultaneously enriched with zinc and n?3 highly unsaturated fatty acids (HUFAs), the zinc content increased, but the n?3 HUFA content did not. Therefore, separate enrichment with zinc and fatty acids was adopted. The zinc content of rotifers fed zinc-enriched Chlorella was significantly higher than that of rotifers fed unenriched Chlorella. After zinc enrichment, rotifers were enriched with fatty acids, and the docosahexaenoic acid (DHA) and n?3 HUFA levels in rotifers were higher than the levels obtained after simultaneous enrichment with zinc and fatty acids. With regard to the concentration of other minerals in rotifers after zinc enrichment, the manganese content tended to decrease when the zinc content increased.The results of this study demonstrated that zinc enrichment of rotifers was successfully performed by using microalgae that had accumulated zinc, and the enrichment of rotifers with fatty acids was also achieved after the completion of zinc enrichment and before feeding the larvae. This method could be utilized for the enrichment of zooplankton with other minerals as well.     

Nutritional value and production of three species of purple non‐sulphur bacteria grown in palm oil mill effluent and their application in rotifer culture

Three species of purple non‐sulphur bacteria (PB), Rhodopseudomonas palustris, Rhodobacter sphaeroides and Rhodovulum sulfidophilum, grown in palm oil mill effluent (POME) were successfully used for the first time as feed for rotifers (Brachionus rotundiformis). Rp. palustris cultured in both POME and synthetic medium gave the highest rotifer density (332–395 individuals mL⁻¹) from 3 to 5 days at 10 g L⁻¹ salinity. Other PB cultured in synthetic medium generally support higher rotifer density than PB cultured in POME. Rb. sphaeroides had the highest biomass (1.91–3.34 g L⁻¹) and growth rate (0.64–1.11 g day⁻¹) in both types of culture medium. Nevertheless, only Rv. sulfidophilum grown in POME contained both eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), indicating its ability to biosynthesize them from POME nutrients. Rotifers fed Rv. sulfidophilum grown in POME had significantly higher amounts of protein, arachidonic acid, EPA and DHA than rotifers fed Rv. sulfidophilum grown in synthetic medium. The nutritional profile of lipid‐deficient PB can be improved by growing them in POME, and these enriched PB produced at an estimated cost of USD 8.71–35.35 kg⁻¹ dry biomass, depending on species, can support rotifer production in a batch culture system.     

The enrichment of rotifers (Brachionus plicatilis) and Artemia franciscana with taurine liposomes and their subsequent effects on the larval development of California yellowtail (Seriola lalandi)

Taurine is an essential or conditionally essential nutrient for many species of marine fish, especially during early development. There is growing evidence that marine fish larvae benefit from taurine‐enriched rotifers; however, it is unknown if larvae benefit from taurine‐enriched Artemia. We investigated the effects of taurine‐enriched rotifers (Brachionus plicatilis) and Artemia franciscana on the growth and whole‐body taurine concentrations of California yellowtail (Seriola lalandi; CYT) larvae. The approach used in this study was to encapsulate taurine within microparticles (liposomes), which were then fed to rotifers and Artemia. We found that feeding taurine liposomes to rotifers and Artemia resulted in taurine concentrations in these prey species that were similar to or above those previously reported in copepods. At the end of the rotifer phase, CYT larvae fed taurine‐enriched rotifers showed increased growth (final dry weights; DW) and had higher whole body taurine concentrations when compared to larvae fed unenriched rotifers. At the end of the Artemia phase, CYT whole body taurine concentrations varied among dietary treatments. Larval lengths and DWs were not significantly different among treatments at the end of the Artemia phase, suggesting that the taurine concentrations of unenriched Artemia were sufficient to support the growth of CYT larvae.     

Nutrient evaluation of rotifers and zooplankton: feed for marine fish larvae

The objective of this study was to examine the biochemical composition of intensively reared rotifers after enrichment with three commonly used enrichment media, Multigain, Ori‐Green and DHA‐enriched freshwater algae Chlorella, using standard enrichment protocols at a local cod larvae producer and compare it with that of natural zooplankton from Lofilab AS, a cod larvae producer using semi‐intensive rearing techniques. Unenriched rotifers were analysed to examine whether the enrichment procedures were successful in increasing the content of essential nutrients to level requirements for marine fish larvae. Neither total lipids nor proteins were affected by enrichment. Unenriched rotifers were significantly lower in highly unsaturated fatty acids (HUFAs) and significantly higher in linoleic acid (LA, C18:2, n‐6), than were zooplankton. Enrichment with Chlorella and Multigain increased the HUFAs significantly, while they were slightly reduced after enrichment with Ori‐Green. Total amino acids and mineral content were unaffected by enrichment. Zooplankton was rich in taurine and selenium, whereas rotifers were devoid of it, both prior to and after enrichment. Using zooplankton as a reference for the nutritional requirements of marine fish larvae, results from this study demonstrate that enrichment media currently in use are not effective for enhancing the nutritional quality of rotifers.     

The effect of dietary n-3 HUFA levels and DHA/EPA ratios on growth, survival and osmotic stress tolerance of Chinese mitten crab Eriocheir sinensis larvae

The effect of varying levels of dietary n-3 highly unsaturated fatty acid (HUFA) and docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratios on growth, survival and osmotic stress tolerance of Eriocheir sinensis zoea larvae was studied in two separate experiments. In experiment I, larvae were fed rotifers and Artemia enriched with ICES emulsions with 0, 30 and 50% total n-3 HUFA levels but with the same DHA/EPA ratio of 0.6. In experiment II, larvae were fed different combinations of enriched rotifers and Artemia, in which, rotifers were enriched with emulsions containing 30% total n-3 HUFA, but different DHA/EPA ratio of 0.6, 2 and 4; while Artemia were enriched with the same emulsions, but DHA/EPA ratio of 0.6 and 4. In both experiments, un-enriched rotifers cultured on baker's yeast and newly-hatched Artemia nauplii were used as control diets. Larvae were fed rotifers at zoea 1 and zoea 2 stages; upon reaching zoea 3 stage, Artemia was introduced.Experiment I revealed no significant effect of prey enrichment on the survival of megalopa among treatments, but higher total n-3 HUFA levels significantly enhanced larval development (larval stage index, LSI) and resulted in higher individual dry body weight of megalopa. Furthermore higher dietary n-3 HUFA levels also resulted in better tolerance to salinity stress. Experiment II indicated that at the same total n-3 HUFA level, larvae continuously receiving a low dietary DHA/EPA ratio had significantly lower survival at the megalopa stage and inferior individual body weight at the megalopa stage, but no negative effect was observed on larval development (LSI). The ability to endure salinity stress of zoea 3, zoea 5 and megalopa fed diets with higher DHA/EPA ratio was also improved.     

Effect of different enrichment products rich in docosahexaenoic acid on growth and survival of meagre,Argyrosomus regius (Asso, 1801)

Live prey used in aquaculture to feed marine larval fish – rotifer and Artemia nauplii – lack the necessary levels of n‐3 polyunsaturated fatty acids (n‐3 PUFA) which are considered essential for the development of fish larvae. Due to the high voracity, visual feeding in conditions of relatively high luminosity, and cannibalism observed in meagre larvae, a study of its nutritional requirements is needed. In this study, the effect of different enrichment products with different docosahexaenoic acid (DHA) concentrations used to enrich rotifers and Artemia metanauplii have been tested on growth, survival, and lipid composition of the larvae of meagre. The larvae fed live prey enriched with Algamac 3050 (AG) showed a significantly higher growth than the rest of the groups at the end of the larval rearing, while the larvae fed preys enriched with Multigain (MG) had a higher survival rate. DHA levels in larvae fed prey enriched with MG were significantly higher than in those fed AG‐enriched prey. High levels of DHA in Artemia metanauplii must be used to achieve optimal growth and survival of meagre larvae.     

Evaluation of nutritional and environmental variables during early larval culture of pigfish Orthopristis chrysoptera (Linnaeus)

Pigfish (Orthopristis chrysoptera Linnaeus) are a commonly used baitfish in the southeastern United States. Aquaculture methods for broodfish spawning and juvenile grow‐out have been developed but there is still a paucity of information regarding larval culture methods. Five, short duration (10 days) experiments were conducted to determine effective strategies to yield high larval survival and growth during early development. Experiment one examined the rotifer enrichments Ori‐Green, DHA Protein Selco, and AlgaMac 3050 as well as a non‐enriched control along with corresponding fatty acid levels in the enriched rotifers and pigfish larvae. Experiment two evaluated three, once daily feeding frequencies of either 5, 10 or 20 rotifers mL^?1. Experiment three compared feeding 20 rotifers mL^?1 once daily to feeding 5 rotifers mL^?1 twice daily. Experiment four examined four different larval stocking densities: 50, 75, 100, or 125 larvae L^?1. Experiment five examined green water strategies using either live Tahitian strain Isochrysis galbana (Parke) or Nannochloropsis oculata (Hibberd) paste at either 250 000 or 500 000 cells mL^?1 as well as a clear water control. Results indicated rotifer enrichment with DHA Protein Selco and green water application using live T‐ISO at 500 000 cells mL^?1 had the highest survival of pigfish during early stages of larval culture. A once daily rotifer feeding regime of 20 rotifers mL^?1 and stocking density of 50 larvae L^?1 also improved survival. These results provide producers with methods to improve efficiency for pigfish larval culture and provide researchers with new foundational data, such as potential fatty acid requirements.     

Relationship between prey utilization and growth variation in hatchery‐reared Pacific bluefin tuna,Thunnus orientalis (Temminck et Schlegel), larvae estimated using nitrogen stable isotope analysis

In mass culture of Pacific bluefin tuna Thunnus orientalis, a marked growth variation is observed after they start feeding at 6–7 mm in body length (BL) on yolk‐sac larvae of other species, and the growth variation in tuna larvae is a factor leading to the prevalence of cannibalism. To examine the relationship between prey utilization and growth variation, nitrogen stable isotope ratios (δ¹⁵N) of individual larvae were analysed. A prey switch experiment was conducted under two different feeding regimes: a group fed rotifers (rotifer fed group), and a group fed yolk‐sac larvae of spangled emperor, Lethrinus nebulosus (fish fed group) from 15 days after hatching (6.87 mm BL). The fish fed group showed significantly higher growth than the rotifer fed group. Changes in the δ¹⁵N of the fish fed group were expressed as an exponential model and showed different patterns from those of the rotifer fed group. The δ¹⁵N of fast‐growing tuna larvae collected in an actual mass culture tank after the feeding of yolk‐sac larvae was significantly higher than those of the slow‐growing larvae, indicating that slow glowing larvae depended largely on rotifers rather than the yolk‐sac larvae.     

Purified deoxynivalenol or feed restriction reduces mortality in rainbow trout,Oncorhynchus mykiss (Walbaum), with experimental bacterial coldwater disease but biologically relevant concentrations of deoxynivalenol do not impair the growth of Flavobacterium psychrophilum

Diets containing deoxynivalenol (DON) were fed to rainbow trout Oncorhynchus mykiss (Walbaum) for 4 weeks followed by experimental infection (intraperitoneal) with Flavobacterium psychrophilum (4.1 × 10⁶ colony‐forming units [CFU] mL⁻¹). Mortality of rainbow trout fed either 6.4 mg kg⁻¹ DON or trout pair‐fed the control diet was significantly reduced (P < 0.05) in comparison with trout fed the control diet to apparent satiation (<0.1 mg kg⁻¹ DON). In a second experiment, trout were fed one of three experimental diets; a control diet, a diet produced with corn naturally contaminated with DON (3.3 mg kg⁻¹ DON) or a diet containing purified DON (3.8 mg kg⁻¹); however, these fish were not experimentally infected. The presence of DON resulted in significant reduction (P < 0.0001) in feed intake as well as weight gain after 4 weeks. Respiratory burst of head‐kidney leucocytes isolated from rainbow trout fed diets containing purified DON (3.8 mg kg⁻¹) was significantly higher (P < 0.05) at 35 day post‐exposure compared with controls. The antimicrobial activity of DON was examined by subjecting F. psychrophilum in vitro to serial dilutions of the chemical. Complete inhibition occurred at a concentration of 75 mg L⁻¹ DON, but no effect was observed below this concentration (0–30 mg L⁻¹).     

Effects of co‐supply ratios of swimming crab Portunus trituberculatus zoeae and Artemia on survival and growth of East Asian common octopus Octopus sinensis paralarvae under an upwelling culture system

Despite recent advances in culture technology for East Asian common octopus Octopus sinensis paralarvae using upwelling systems, securing suitable feed for the paralarvae is an unresolved issue. The zoea of the swimming crab Portunus trituberculatus is a good candidate for paralarval feed because of the high fecundity of the adult females. To investigate the effects of supplying P. trituberculatus zoeae and their feeding method on paralarvae, we cultured paralarvae with supplying different combination ratios of zoeae and Artemia (10:0, 7:3, 5:5, 3:7 and 0:10), and with or without supplementing rotifers using small‐scale (3‐L) upwelling systems. Paralarval survival rate and growth were improved when zoeae were supplied as the main feed, but reduced when the proportion of Artemia exceeded half the whole preys. Supplementing rotifers did not affect the paralarval survival and growth. Subsequently, paralarvae were cultured by supplying zoeae (partially augmented by Artemia) using three large (1‐kl) upwelling systems to assess their feeding effectiveness in juvenile octopus production. Paralarvae could be cultured at high survival rates of 77.1 ± 5.1% to reach benthic juveniles at 23 days after hatching. In conclusion, supplying P. trituberculatus zoeae augmented with Artemia under an upwelling culture system has great potential for juvenile octopus production.