Immunostimulatory effects of dietary poly‐β‐hydroxybutyrate in European sea bass postlarvae

The stable production of high‐quality fry in marine aquaculture is still hampered by unpredictable mortality caused by infectious diseases during larval rearing. Consequently, the development of new biocontrol agents is crucial for a viable aquaculture industry. The bacterial energy storage compound poly‐β‐hydroxybutyrate (PHB) has been shown to exhibit beneficial properties on aquatic organisms such as enhanced survival, growth, disease resistance and a controlling effect on the gastrointestinal microbiota. However, the effect of PHB on the developing immune system of fish larvae has so far not been investigated. In this study, the effect of feeding PHB‐enriched Artemia nauplii on survival, growth and immune response in European sea bass (Dicentrarchus labrax) postlarvae was examined. Amorphous PHB was administered to 28‐day‐old sea bass postlarvae over a period of 10 days. The survival and growth performance were monitored, and the expression of 29 genes involved in immunity, growth, metabolism and stress‐response was measured. While the expression of the insulin‐like growth factor 1 (igf1), an indicator of relative growth, was upregulated in response to feeding PHB, the larval survival and growth performance remained unaffected. After 10 days of PHB treatment, the expression of the antimicrobial peptides dicentracin (dic) and hepcidin (hep) as well as mhc class IIa and mhc class IIb was elevated in the PHB fed postlarvae. This indicates that PHB is capable of stimulating the immune system of fish early life stages, which may be the cause of the increased resistance to diseases and robustness observed in previous studies.     

Determination of poly‐β‐hydroxybutyrate assimilation by postlarval whiteleg shrimp,Litopenaeus vannamei using stable 13C isotope tracing

A preliminary study was conducted to demonstrate fate of the bacterial storage compound poly‐β‐hydroxybutyrate (PHB) once ingested and degraded in vivo in crustaceans. The 2% supplementation of ¹³C‐labeled Ralstonia eutropha DSM545 containing 75% PHB on dry weight in postlarval whiteleg shrimp, Litopenaeus vannamei resulted in consistent enrichment of two fatty acids (containing 14–24 carbons [C] in length ) pentadecanoic acid (15:0) and palmitic acid (16:0) in all lipid fractions. This could indicate that ¹³C signal may not only originate from the PHB but partly from the structural components of the bacterial cell. There was a trend of lower ¹³C enrichment in various lipid fractions of the postlarvae as compared to the diet. These findings may have important implications as to potential of PHB as energy source rather than as building block. There was a rapid transfer of ¹³C in the postlarvae as seen by the significant ¹³C enrichment in the tissues 4 hr after feeding with labeled bacterial cells. Overall, the assimilation of PHB in crustacean tissues is demonstrated for the first time. Our results indicate that PHB once ingested is rapidly assimilated in the tissues and could probably act as an energy source rather than as a building block.     

Zingiber officinalis an herbal appetizer in the tiger shrimp Penaeus monodon (Fabricius) larviculture

Penaeus monodon postlarvae were fed with different percentages (0%, 25%, 50%, 75% and 100%) of the herbal appetizer Zingiber officinalis enriched Artemia. After 30 days of culture (i.e. PL‐1–30), a very positive result was found in Z. officinalis‐enriched Artemia‐fed postlarvae. The unenriched Artemia‐fed postlarvae consumed 91.0 mg/animal/30 days of feed, whereas the Z. officinalis‐enriched Artemia increased their consumption to 127.9 mg/animal/30 days. A similar pattern was noticed in feed absorbed (110.2 mg), dry weight growth (26.7 mg) and feed catabolized (83.2 mg) in Z. officinalis‐enriched Artemia because of enzymatic activities. The conversion efficiency of unenriched postlarva was 17.19%, whereas in 100%Z. officinalis‐enriched Artemia, the maximum conversion efficiency was 20.85%. The net production efficiency increased significantly (P < 0.05) to 22% from that of the unenriched Artemia‐fed postlarvae. The administration of Z. officinalis in all levels produced significantly (P < 0.05) higher weight gain and specific growth rate. The utilization efficiency of feed increased proportionately to the percentages of Z. officinalis. Digestive enzyme activity (amylase, protease and lipase) increased significantly (P < 0.05) in the 50%, 75% and 100% enrichment. Among the different percentages of enrichment, the 100%Z. officinalis‐enriched Artemia‐fed postlarvae performed better in the overall status.     

Change in nutrient composition of Artemia grown for 3–4 days and effects of feeding on‐grown Artemia on performance of Atlantic halibut (Hippoglossus hippoglossus,L.) larvae

Major challenges in culture of Atlantic halibut larvae have been slow growth during the late larval stages and inferior juvenile quality due to pigmentation errors and incomplete eye migration during metamorphosis. The hypothesis of this study was that feeding on‐grown Artemia would alleviate these problems. Artemia were grown for 3–4 days on Origreen or Origo. The growth and nutrient composition of Artemia nauplii and on‐grown Artemia were analysed, and both Artemia types were fed to Atlantic halibut larvae, on‐grown Artemia from 15 days post‐first feeding (dpff). The body length of Artemia increased with 20%–70% in response to on‐growing. In all experiments, protein, free amino acids and the ratio of phospholipid to total lipid increased, while lipid and glycogen decreased. The fatty acid composition improved in some cases and not in others. The micronutrient profiles were not negatively affected in on‐grown Artemia. All these changes are thought to be beneficial for marine fish larvae. The final weight of Atlantic halibut postlarvae was similar, and 90% of the juveniles had complete eye migration in both groups. It is concluded that the present version of Artemia nauplii probably covers the nutrient requirements of Atlantic halibut larvae.     

Siberian sturgeon (Acipenser baerii) larvae fed Artemia nauplii enriched with poly‐β‐hydroxybutyrate (PHB): effect on growth performance,body composition,digestive enzymes,gut microbial community,gut histology and stress tests

The effects of feeding Artemia nauplii enriched with or without poly‐β‐hydroxybutyrate (PHB) and/or highly unsaturated fatty acids (HUFA) on Siberian sturgeon (Acipenser baerii) larvae were investigated. Feeding larvae with PHB‐enriched nauplii (PHB, PHB + HUFA) decreased the growth performance of the larvae. PHB affected the body composition by increasing the lipid content of the whole body and decreasing total saturated, monoenoic, n3, n6 and decosahexanoeic acid (DHA) in the larvae, indicating that the PHB addition affected lipid metabolism. A high activity of pepsin was observed in the digestive extracts of PHB treatments (PHB, PHB + HUFA), while PHB suppressed amylase activity in the intestine of the larvae. Based on molecular analysis, PHB changed the microbial community in the distal intestine of the larvae. The highest counts of goblet cells were observed in the HUFA‐containing treatments (HUFA, PHB + HUFA), indicating that HUFA addition may improve the mucosal barrier defence system. The overall quality of the larvae was evaluated by exposing them to different salinities and ammonia stress levels. PHB decreased survival rates in these challenges. Our results show that optimal PHB doses for bio‐encapsulation into Artemia remain to be determined for further application at the earliest larval stages of sturgeon.     

Artificial substratum consisting of poly‐β‐hydroxybutyrate‐based biodegradable plastic improved the survival and overall performance of postlarval tiger shrimp Penaeus monodon

The use of artificial substratum consisting of poly‐β‐hydroxybutyrate (PHB)‐based biodegradable plastic for penaeid shrimp culture was investigated in the present study. The survival of postlarval tiger shrimp Penaeus monodon (30 ± 5 mg) provided with PHB substratum made out of PHB type DP9002 (Metabolix GmbH, Köln, Germany) was 88.7 ± 3.4% and this was significantly higher as compared to postlarvae provided conventional substratum consisting of polyvinylchloride (PVC) pipes (67.3 ± 6.5%). However, no significant weight improvement was observed for the postlarval tiger shrimp indicating that PHB could not be used as growth promoter. Nevertheless, a trend of improved robustness against adverse environmental conditions (lethal ammonium chloride concentration) and increased resistance to pathogenic Vibrio was observed in postlarval tiger shrimp provided with PHB substratum as compared to postlarvae provided with PVC substratum. Results indicate higher preference by postlarvae on PHB substratum over PVC substratum. Overall, this study indicates the potential of artificial substratum consisting of PHB‐based biodegradable plastic as replacement for conventional substratum consisting of PVC pipes in enhancing the survival of postlarval tiger shrimp and improving its performance against adverse environmental conditions and disease resistance.     

Antioxidant activity and lipid peroxidation in Artemia nauplii enriched with DHA‐rich oil emulsion and the effect of adding an external antioxidant based on hydroxytyrosol

Artemia nauplii catabolize polyunsaturated fatty acids (PUFA); in particular, they retroconvert docosahexaenoic acid (DHA, 22:6n‐3), so enrichment is a continuous quest towards increasing PUFA through the use of PUFA‐rich enrichment products. However, optimal conditions during enrichment (aeration, illumination and temperatures around 28°C) tend to accelerate autoxidation of PUFA and the formation of potentially toxic oxidation products. Water‐soluble antioxidants like the polyphenolic compound hydroxytyrosol (3,4‐dihydroxyphenylethanol), a polar molecule found in the water fraction resulting after the milling process of olives, arise as promising compounds to prevent oxidation during Artemia enrichments. We investigated the antioxidant activity and lipid peroxidation in Artemia nauplii during enrichment and the effect of adding an external antioxidant based on hydroxytyrosol during the enrichment with a PUFA‐rich emulsion (M70). For this purpose, the activity of antioxidant enzymes (catalase, superoxide dismutase, glutathione‐S‐transferase, glutathione peroxidase), as well as lipid peroxidation, was determined in enriched and unenriched Artemia nauplii. To validate antioxidant activity and lipid peroxidation, in a first experiment, nauplii were enriched with microalgae (Tetraselmis suecica), yeast (Saccharomyces cerevisiae) and M70 emulsion. In a second experiment, enrichment with a commercial emulsion (DC Super Selco), M70, and a combination of M70 and hydroxytyrosol (Hytolive, HYT) added as an external antioxidant were performed. The combination of M70 with HYT produced the best results, in terms of activity of antioxidant enzymes. The analysis of the fatty acids from total lipids showed that the addition of hydroxytyrosol preserved the DHA percentage of enriched nauplii.     

Effect of hot water extracts of brown seaweeds Sargassum spp. on growth and resistance to white spot syndrome virus in shrimp Penaeus monodon postlarvae

An experiment was conducted to evaluate the effect of a hot water extract of brown seaweeds Sargassum duplicatum and Sargassum wightii on the growth and white spot syndrome virus (WSSV) resistance in shrimp Penaeus monodon postlarvae (PL). Artemia nauplii (instar II) were enriched with both seaweed extracts at various concentrations (250, 500 and 750 mg L^?1) and fed to the respective P. monodon (PL15–35) group for 20 days. A control group was also maintained without seaweed extract supplementation. The weight gain of the experimental groups was significantly higher (0.274–0.323 g) than the control group (0.261 g). Similarly, the specific growth rate was also significantly higher (16.27–17.06%) in the experimental groups than in the control group (16.03%). After 20 days of the feeding experiment, the shrimp PL were challenged with WSSV for 21 days. During the challenge test, the control shrimp displayed 100% mortality within 8 days. In contrast, the mortality percentage of the highest concentration (750 mg L^?1) of seaweed extract enriched Artemia nauplii fed shrimp was 54–79%. Comparatively, low mortality was observed in S. wightii extract‐enriched Artemia nauplii fed shrimp. The polymerase chain reaction analysis indicated the concentration‐dependent infection of WSSV in P. monodon PL.     

Survival and Growth Improvement of Palm Ruff,Seriolella violacea,Larvae Fed Artemia Nauplii Enriched with an Experimental Emulsion

The palm ruff, Seriolella violacea (Cojinoba), is a potential new species for Chilean aquaculture. To approach Cojinoba larviculture, an experimental Artemia enrichment emulsion, containing docosahexaenoic acid (DHA)/eicosapentaenoic acid (EPA) = 2.5, supplemented with vitamin E, astaxanthin, and β‐glucan, was evaluated in both Artemia and Cojinoba larvae, 30–50 d.a.h. This study tested an experimental enrichment emulsion versus a commercial emulsion, with an integral approach of multicompound emulsions. After 23 h enrichment, experimental emulsion (EE)‐enriched nauplii reached DHA and EPA concentrations of 23.8 and 18.7 mg/g dry weight (dwt), respectively, while in Cojinoba larvae they were 18.4 and 19.7 mg/g dwt. Control emulsion (CE)‐enriched nauplii exhibited lower DHA and EPA (6.1 and 7.7 mg/g dwt), while only DHA decreased in the control larvae (12.6 mg/g dwt). Vitamin E was higher in EE‐enriched nauplii (29.2 mg/100 g dwt) than in the control (8.4 mg/100 g dwt). Larvae fed EE‐enriched Artemia exhibited 8% increase in survival and 19% in growth compared with the control. Astaxanthin was detected only in larvae fed EE‐enriched nauplii. The tumor necrosis factor‐α concentration was not significantly different between larvae fed EE‐ and CE‐enriched nauplii. EE looks promising as an Artemia enrichment and experimental diet to assess palm ruff larval requirements, and has a positive impact on fish larvae performance.     

Enrichment of Artemia metanauplii in phospholipids and essential fatty acids as a diet for common octopus (Octopus vulgaris) paralarvae

Highly unsaturated fatty acids (HUFA), like the eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids and polar lipids (essentially phospholipids, PL) have been identified as essential nutrients for common octopus (Octopus vulgaris) paralarvae. However, they are not available in sufficient amounts in live preys as Artemia, making necessary a supplementation of these nutrients previous use. A commercial emulsion, soya liposomes, and marine and soya lecithins were used to supply HUFA and PL to Artemia metanauplii, those being regarded as suitable size preys for octopus paralarvae. Our results prove that a simultaneous enrichment in HUFA and PL is possible using enrichment diets combining HUFA‐ and PL‐rich products in short‐term (4 h) incubations. Particularly interesting was the enrichment efficiency shown by the marine lecithin, which enabled the enhancement of the PL fraction of Artemia metanauplii and, importantly, also their HUFA with a remarkable 13% DHA of total fatty acids. Marine lecithin arises as a novel enrichment diet for Artemia and more effective than some commercial products currently used in hatcheries worldwide.     

The performance of larval Seriola lalandi (Valenciennes, 1833) is affected by the taurine content of the Artemia on which they are fed

This study describes the effects of feeding taurine‐supplemented Artemia on the growth, survival, whole body taurine content and jaw malformation rate of larval yellowtail kingfish Seriola lalandi. Larvae were fed rotifers containing no supplemental taurine from 3 to 15 day post hatch (dph) and Artemia co‐enriched with taurine from 12 to 22 dph. Artemia were supplemented at concentrations of either 0, 0.8, 1.6, 2.4, 3.2 or 4.0 g of taurine L^?1 during the 18 h HUFA enrichment process. Taurine content in the Artemia increased from 0.76 ± 0.04% DW in those without supplementation to 3.95 ± 0.17% DW in those supplemented at 4.0 g L^?1. Survival rates of larval yellowtail kingfish were significantly lower in all taurine‐supplemented treatments compared to the unsupplemented control. Growth was significantly improved in those larvae fed taurine‐supplemented Artemia; however, we cannot attribute this improvement solely to taurine, as improved growth may have been a function of the reduced survival, and therefore increased prey availability, in these treatments. The whole body taurine content of larvae fed unsupplemented Artemia was significantly lower (1.85 ± 0.03% DW) than those fed supplemented Artemia, which did not differ from each other (pooled average 2.48 ± 0.03% DW), suggesting either a functional excretion mechanism is in place or that this represents the saturation value for larvae of this age. Jaw malformation rates were not affected by Artemia taurine content. The results of this research suggest yellowtail kingfish larvae may have a lower requirement and/or a reduced tolerance to excess dietary taurine than juveniles.     

Penaeus monodon larvae can be protected from Vibrio harveyi infection by pre‐emptive treatment of a rearing system with antagonistic or non‐antagonistic bacterial probiotics

This study shows that the disease resistance and survival rate of Penaeus monodon in a larval rearing systems can be enhanced by supplementing with antagonistic or non‐antagonistic probiotics. The antagonistic mode of action of Pseudomonas MCCB 102 and MCCB 103 against vibrios was demonstrated in larval mesocosm with cultures having sufficient concentration of antagonistic compounds in their culture supernatant. Investigations on the antagonistic properties of Bacillus MCCB 101, Pseudomonas MCCB 102 and MCCB 103 and Arthrobacter MCCB 104 against Vibrio harveyi MCCB 111 under in vitro conditions revealed that Pseudomonas MCCB 102 and MCCB 103 were inhibitory to the pathogen. These inhibitory properties were further confirmed in the larval rearing systems of P. monodon. All these four probionts significantly improved larval survival in long‐term treatments as well as when challenged with a pathogenic strain of V. harveyi MCCB 111. We could demonstrate that Pseudomonas MCCB 102 and MCCB 103 accorded disease resistance and a higher survival rate in P. monodon larval rearing systems through active antagonism of vibrios, whereas Bacillus MCCB 101 and Arthrobacter MCCB 104 functioned as probiotics through immunostimulatory and digestive enzyme‐supporting modes of action.     

Delivery of HUFA, probionts and biomedicine through bioencapsulated Artemia as a means to enhance the growth and survival and reduce the pathogenesity in shrimp Penaeus monodon postlarvae

One of the major problems in the shrimp culture industry is the difficulty in producing high-quality shrimp larvae. In larviculture, quality feeds containing a high content of highly unsaturated fatty acids (HUFA) and ingredients that stimulate stress and disease resistance are essential to produce healthy shrimp larvae. In the present study, Penaeus monodon postlarvae (PL15) were fed for 25 days on an unenriched Artemia diet (control; A) or on a diet of Artemia enriched with either HUFA-rich liver oil of the trash fish Odonus niger (B), probionts [Lactobacillus acidophilus (C1) or yeast-Saccharomyces cerevisiae (C2)] or biomedicinal herbal products (D) that have anti-stress, growth-promoting and anti-microbial characteristics. P. monodon postlarvae fed unenriched Artemia exhibited the lowest weight gain (227.9 ± 8.30 mg) and specific growth rate (9.95 ± 0.05%), while those fed the HUFA-enriched Artemia (B) exhibited the highest weight gain and specific growth rate (362.34 ± 12.56 mg and 11.77 ± 0.08%, respectively). At the end of the 25-day rearing experiment, the shrimp postlarvae (PL40) were subjected to a salinity stress study. At both low and high (0 and 50‰) salinities, the group fed the control diet (A) experienced the highest cumulative mortality indices (CMI) 935.7 ± 2.1 and 1270.7 ± 3.1, respectively. Those fed diet D showed the lowest stress-induced mortality, and CMI were reduced by 31.1 and 32.3% under conditions of low and high salinity stress, respectively. A 10-day disease challenge test was conducted with the P. monodon postlarvae (PL40–PL50) by inoculating the shrimp with the pathogen Vibrio harveyi at the rate of 10⁵–10⁷ CFU/ml in all rearing tanks. P. monodon postlarvae fed probiont-encapsulated Artemia diets (C1 and C2) exhibited the highest survival (94.3 and 82.3%, respectively) and lowest pathogen load (V. harveyi) in hepatopancreas (5.2 × 10² ± 9.0 × 10 and 4.6 × 10² ± 9.0 × 10 CFU g⁻¹, respectively) and muscle (2.0 × 10² ± 6 × 10 and 1.7 × 10² ± 8.6 × 10 CFU g⁻¹, respectively) tissues. The shrimp that were fed the unenriched Artemia (Control; A) showed the lowest survival (26.33%) and highest bacterial load in the hepatopancreas (1.0 × 10⁵ ± 5 × 10³ CFU g⁻¹) and muscle (3.6 × 10⁴ ± 6 × 10² CFU g⁻¹). The shrimp fed the herbal product (D)-enriched Artemia also exhibited enhanced survival and reduced V. harveyi load in the tissues tested compared to the control diet (A) group. The results are discussed in terms of developing a quality larval feed to produce healthy shrimp larvae.     

Effect of tank colour on larval performance of the Amazon River prawn Macrobrachium amazonicum

The aim of this study was to evaluate the effects of hatchery‐tank colours (white, yellow, red, blue, green and black) on the performance of larval culture of Macrobrachium amazonicum. The larvae were fed daily with newly hatched Artemia nauplii. The hatchery‐tank colours affected the light level inside the tanks, the consumption of Artemia nauplii (AN), larval development, survival, mass gain and productivity of postlarvae (PL). The overall consumption of Artemia nauplii per larva during the larval cycle was 30% and 45% higher in the green and red tanks respectively. The significant variation of AN consumption among tank colours (P = 0.0006) indicates that M. amazonicum larvae are visual predators. Survival was higher in the black, blue and green tanks, reaching more than 75%. However, the highest productivity was obtained in the black tanks (80.1 PL L^?1). Lighter coloured tanks and excess luminosity (more than 2 μmol s^?1 m² at tank bottom) appear to be important stress factors for larvae, contributing to reduce survival and productivity. The results indicate that rearing M. amazonicum in black tanks will improve larvae condition, ensure greater productivity of postlarvae and lower Artemia consumption, increasing technological and economic viability.     

Establishing larval feeding regimens for the Forktail Blenny Meiacanthus atrodorsalis (Günther, 1877): effects of Artemia strain,time of prey switch and co‐feeding period

This study aimed to establish feeding strategies covering the whole larval period of the forktail blenny, Meiacanthus atrodorsalis, based on the standard hatchery feeds of rotifers and Artemia. Three purposely designed experiments were conducted to determine the appropriate times and techniques to transition larvae from rotifers onto Artemia nauplii of a Great Salt Lake (GSL) strain, and a specialty AF strain, as well as subsequent transition onto enriched metanauplii of GSL Artemia. With a 3‐day co‐feeding period, larvae adapted well to a transition from rotifers to newly hatched GSL Artemia nauplii as early as 5 days posthatching (DPH), and as early as 3 DPH when fed the smaller AF Artemia nauplii. However, prolonging the rotifer‐feeding period up to 11 DPH did not negatively affect survival. Larvae fed Artemia nauplii of the AF strain showed 17–21% higher survival, 24–33% greater standard length and body depth, and 91–200% greater dry weight, after 20 days relative to those fed nauplii of the GSL strain. Meanwhile, enriched Artemia metanauplii of the GSL strain were shown to be an acceptable alternative to AF Artemia nauplii for later larvae, producing similar survival and growth when introduced from 8 DPH. Based on our findings, we recommend feeding M. atrodorsalis larvae rotifers as a first food between 0 and 2 DPH, introducing AF Artemia nauplii from 3 DPH, followed by enriched GSL Artemia metanauplii from 8 DPH onward, with a 3‐day co‐feeding period between each prey change.     

Enriching Artemia nauplii with a high DHA‐containing lipid emulsion: search for an optimal protocol

This study aimed to investigate practical strategies to optimize the use of a high‐docosahexaenoic acid (DHA) lipid emulsion (M70), a product with great potential in live prey enrichments for marine larviculture. Considering its particularly high content in DHA (22:6n‐3), the adequate utilization of the emulsion for Artemia enrichments was evaluated in a series of six experiments. More specifically, the bioencapsulation efficiency of M70 into Artemia nauplii was tested under different experimental conditions of oxygen source, aeration flow, incubation temperature, concentration and dosage, as well as nauplial densities. Our results showed that an optimal utilization of M70 is achieved with incubation temperatures of 28°C, moderate aeration flows and nauplial densities of 300 ind per mL. In addition, the emulsion can be dispensed in the enrichment medium in one single dose of 0.8 g L^?1, with no apparent detrimental effects on its oxidative stability and Artemia nauplii survival during enrichment.     

Effects of dietary astaxanthin supplementation on survival,growth and stress resistance in larval and post‐larval kuruma shrimp,Marsupenaeus japonicus

Two trials were conducted to evaluate the effects of chemically synthesized astaxanthin (Ax) on growth performance, survival, and stress resistance in larval and post‐larval kuruma shrimp (Marsupenaeus japonicus). Six dietary levels of Ax (0, 50, 100, 200, 400 and 800 mg/kg diet) were added to a baseline diet. As a first study, an 8‐day feeding trial was conducted on larval kuruma shrimp, with results showing that larvae‐fed diets supplemented with different Ax levels exhibited better performance during developmental and metamorphosis to postlarvae. Broken‐line regression analysis indicated that the optimal levels for growth and stress resistance of larvae were 168.9 mg/kg and 82.1 mg/kg diet, respectively. Second, a 30‐day feeding trial was conducted on post‐larval shrimp. Supplementation with 100 and 200 mg/kg Ax yielded significantly higher final body weight, body weight gain and specific growth rate than that in a control group. A cumulative mortality index for osmotic stress revealed significantly better performance in a group fed 200 mg/kg supplementation than in controls. Broken‐line regression analysis indicated that the optimal levels for growth and stress resistance of postlarvae were 108.7 mg/kg and 178.1 mg/kg diet, respectively. Based on these results, we suggest that carefully dosed Ax supplementation is a beneficial nutritional strategy for the early developmental stages of kuruma shrimp.     

Effect of Diet Processing Method and Ingredient Substitution on Feed Characteristics and Survival of Larval Walleye, Sander vitreus

Two methods were developed for the production of larval fish diets. The first method, microextrusion marumerization (MEM), has been tested in laboratory feeding trials for many years and produces particles that are palatable and water stable. The second method, particle‐assisted rotational agglomeration (PARA), produced diets that have lower density than diets produced by MEM. Each method was used to produce diets in the 250‐ to 400‐ and 400‐ to 700‐μm range and compared with a reference diet (Fry Feed Kyowa * [FFK]) for feeding larval walleye in two experiments. The effect of substituting 4% of the fish meal with freeze‐dried artemia fines was also investigated. In the first experiment, 30‐d survival was greater (P < 0.05) for fish fed a diet produced by PARA without Artemia (49.1.0%) than for fish fed the same diet produced by MEM (27.6%). The addition of Artemia to a diet produced by MEM did not increase survival of larval walleye. Fish fed the reference diet had 24.4% survival. In the second experiment, there was an effect of both processing method and Artemia supplementation, and an interaction of these effects, on survival. Fish fed a diet produced by PARA without Artemia supplementation had 48.4% survival, and fish fed the same diet produced by MEM had only 19.6% survival. Inclusion of 4% freeze‐dried Artemia improved (P < 0.04) survival of fish fed MEM particles but not those fed PARA particles. Fish fed FFK had greater weight gain than fish fed other diets in both experiments. Data indicate that the PARA method of diet processing produces smaller, lower density particles than the MEM process and that diets produced by the PARA process support higher survival of larval walleye with low capital and operating costs.     

Effect of poly‐β‐hydroxybutyrate on growth,enzyme activity and intestinal microbial community of Chinese mitten crab,Eriocheir sinensis (Milne‐Edwards) juveniles

Poly‐β‐hydroxybutyrate (PHB) is microbial carbon and energy storage polymer, which can be degraded into water‐soluble β‐hydroxybutyric acid in the gastrointestinal tract of aquatic animals. A 60‐day culture experiment was performed using Chinese mitten crab, Eriocheir sinensis (Milne‐Edwards) juveniles with an average initial body weight of 0.74 ± 0.06 g which were fed a diet supplemented with 0%, 0.5%, 1%, 3% or 5% PHB. A PHB dietary supplementation of 1% and 3% significantly improved the body weight gain, moulting frequency and concomitantly reduced 2nd–3rd moulting intervals of the crabs (P < 0.05). The dietary PHB level positively related to hepatopancreatic pepsin, trypsin and lipase activity (P < 0.05). Increasing the dietary PHB also improved total superoxide dismutase activity, but reduced alkaline phosphatase and acid phosphatase activity in the serum of hemolymph (P < 0.05). A 16S rRNA gene analysis by denaturing gradient gel electrophoresis indicated that PHB supplementation led to a significantly higher range‐weighted richness, diversity and evenness of the gut bacterial community when dosed at 3% in the feed. The beneficial effects of PHB are discussed in terms of immune defense, metabolism and gut microbiota of the crabs.     

Antiviral Effect of Fucoidan Extracted from the Brown Seaweed,Sargassum wightii,on Shrimp Penaeus monodon Postlarvae against White Spot Syndrome Virus

This study was carried out to determine the effect of seaweed polysaccharide, fucoidan from brown seaweed, Sargassum wightii, on Penaeus monodon postlarvae (PL) against white spot syndrome virus (WSSV). The fucoidan was extracted from S. wightii, and the yield was observed as 2.832 ± 0.204%. Artemia franciscana nauplii were enriched with extracted fucoidan at four different concentrations (100, 200, 300, and 400 mg/L) and fed to shrimp P. monodon PL for 20 d. After feeding experiment, the P. monodon PL were challenged with WSSV, and the mortality percentage was recorded daily up to 21 d. During the challenge test, the control PL showed 100% cumulative mortality within 9 d, but the fucoidan‐enriched Artemia nauplii fed groups of PL exhibited 60–94% cumulative mortality within 21 d. The reduction in mortality percentage of experimental groups of PL over control PL was ranged between 33.71 and 61.65%. The polymerase chain reaction analysis confirmed the concentration‐dependent variation in WSSV infection.