Different responses between orange variant and cultured population of the Pacific oyster Crassostrea gigas at early life stage to temperature‐salinity combinations

Although breeding of rare shell colour variants has drawn widespread attention from shellfish breeders, the potential disadvantages of their adaptive capacity have been ignored in practice. To explore the difference in adaptive capacity between orange shell variant (OSO) and commercially cultured population (CPO) of the Pacific oyster Crassostrea gigas at early life stage, the development to D‐larvae and larval survival and growth (just 23 and 30°C for larval experiment) of them were compared under different temperature (16, 23 and 30°C) and salinity (17, 25 and 33 psu) combinations. In this study, at 23°C and 25 psu, for both OSO and CPO there was no difference in fertilization rates and survival (p > .05) (mean percentages of D‐larvae after fertilized 40 hr ≥ 95.00%; mean larval survival rates on day 10 > 80.00%). However, the percentage of D‐larvae of CPO at 40 hr was significantly (p < .05) higher than OSO at temperatures of 16 and 30°C and 25–33 psu and 17 psu at 23°C. Similarly, CPO has a better larval survival on day 10 and growth than OSO at salinities of 17 and 33 psu at 23°C. Overall, our results indicate that OSO can have an equally good performance like CPO at early life stage under optimal condition (23°C; 25 psu), but the potential disadvantages in adaptive capacity will be shown at suboptimal conditions. These findings can guide future hatchery breeding of OSO, and suggest the potential disadvantages in adaptive capacity in rare colour variants need more attention in further breeding.     

Quality evaluation of indoor‐ and outdoor‐cultured sea cucumber (Apostichopus japonicus) seedlings: Insight from survival and immune performance in response to combined stress of hyperthermia and hyposalinity

The mass mortalities of sea cucumber Apostichpous japonicus have prevailed in northern China, mainly attributing to the emergence of extreme environmental conditions, that is hyperthermia and hyposalinity. The high‐quality sea cucumber seedlings appear to possess more robust resistance to adverse conditions. There are usually indoor‐ and outdoor‐cultured seedlings in industrial production of sea cucumbers. Although the outdoor‐cultured sea cucumbers are practically considered to be more strong and robust, the effective evaluation approach to distinguish these seedlings has been scarce. The current study compared survival and immune performances of indoor‐ and outdoor‐cultured A. japonicus under combined exogenous stressors, that is hyperthermia and hyposalinity. Results based on secondary stress induction revealed that the activities of immune enzymes and levels of catecholamines in body wall of outdoor‐cultured seedlings were prominently higher than those of indoor‐cultured seedlings recovered for 0–72 hr following sublethal stress (30°C and 25 psu of salinity). The opposite case occurred on immune enzymes in coelomic fluid of the two sources of seedlings except for myeloperoxidase. Importantly, the outdoor‐cultured seedlings, which were recovered for 72 hr after sublethal stress, exhibited a 93% of cumulative survival rate following 7 days of recovery after lethal stress (33°C and 20 psu of salinity), 27% higher than the indoor‐cultured seedlings. Collectively, the outdoor‐cultured A. japonicus seedlings showed more superior quality than the hatchery‐produced seedlings in terms of survival and immune performance. These findings provide practically useful information towards quality distinction of the indoor‐ and outdoor‐cultured sea cucumbers, which could benefit the aquaculture industry to obtain high‐quality seedlings.     

Effect of dietary lipid level on growth,lipid metabolism and health status of the Pacific white shrimp Litopenaeus vannamei at two salinities

Three isonitrogenous diets containing 60 g kg^–1, 90 g kg^–1 or 120 g kg^–1 lipid were formulated and fed to the Litopenaeus vannamei (2.00 ± 0.08 g) under two salinities (25 or 3 psu) in triplicate for 8 weeks. Shrimp fed 90 g kg^–1 lipid had higher weight gain and specific growth rate than shrimp fed the other two diets regardless of salinity, and the hepatosomatic index increased with increasing dietary lipid at both salinities. The shrimp at 3 psu had significantly lower survival and ash content, higher condition factor, weight gain and specific growth rate than the shrimp at 25 psu. Increasing dietary lipid level induced the accumulation of serum MDA regardless of salinity, and at 3 psu, it reduced the serum GOT and GPT activities and the mRNA expression of TNF‐α in intestine and gill of L. vannamei. The hepatopancreatic triacylglycerol lipase (TGL) and CPT‐1 mRNA expression showed the highest value in shrimp fed 90 g kg^–1 lipid diet at 3 psu. This study indicates that 120 g kg^–1 dietary lipid may negatively affect the growth and induce oxidative damage in shrimp, but can improve immune defence at low salinity; 60 g kg^–1 dietary lipid cannot afford the growth and either has no positive impact on the immunology for L. vannamei at 3 psu.     

Fluctuating salinity effect on Sphoeroides annulatus (Jenyns 1842) physiological responses

Euryhaline fish, such as the Bullseye puffer Sphoeroides annulatus (Jenyns 1842), experience sudden salinity changes in their natural environment, which is more common than the exception, so they must adapt to survive and cope with extreme salt conditions. Therefore, Bullseye puffer juveniles were exposed to short‐term stress (39 hr) by fluctuating salinity conditions (41, 35, 29, 23, 17, 11, 5, 11, 17, 23, 29, 35, 41 psu) with a 3‐hr interval between each point at 26 ± 1ºC in a respirometer chamber and acclimation reservoirs. Responses to oxygen consumption rate (OCR: 23–35 mg O₂ h^–1 kg^–1), ammonium excretion rate (AER: 1–1.85 mg NH4⁺ h^?1 kg^?1), oxygen‐nitrogen atomic ratio (O:N 17–30), osmoregulatory pattern (blood osmotic pressure from 342.4 to 332.8 mmol/kg) and changes in expression levels of Na⁺/K⁺‐ATPase in the gills (higher values at higher salinities) were measured. Although some signs of stress were detected below the iso‐osmotic point (11.4 psu), the puffer fish is a strong euryhaline fish that survives under these conditions. Nonetheless, it could recover when salinity returned to the initial acclimation point because Sphoeroides annulatus is able to live in a wide range of environments with wide natural salinity fluctuations; thus, a common practice in aquaculture has been to expose fish to low salinity for several reasons discussed in this study. This capacity reveals its high plasticity to saline adaptation from 41 to 5 psu an up from 5 to 41 psu, all in less than 2 days.     

Hypoxia‐induced changes in survival,immune response and antioxidant status of the Pacific white shrimp (Litopenaeus vannamei) fed with graded levels of dietary myo‐inositol

A 3‐hr experiment was conducted to investigate the effects of dietary myo‐inositol (MI) supplementation on survival, immune response and antioxidant abilities in Litopenaeus vannamei under acute hypoxia stress. Six practical diets were formulated with supplementation of graded levels (control group 0, 0.1, 0.2, 0.4, 0.8 and 1.6 g/kg dry diet) of MI and were randomly assigned to triplicate groups of L. vannamei (mean weight 0.40 ± 0.00 g) for 8 weeks. Ten healthy shrimp (final mean weight approximately 11–14 g) randomly selected from each tank were exposed to hypoxia stress after feeding trial. After 3‐hr acute hypoxia stress, survival of shrimp fed MI‐supplemented diets (except 0.1 and 0.4 g/kg diets) was significantly increased compared with the control group. Shrimp fed control diet had lower activities of alkaline phosphatase (AKP), acid phosphatase (ACP), total antioxidant capacity (T‐AOC) and glutathione peroxidase (GPX), and higher malondialdehyde (MDA) and protein carbonyl (PC) contents in hepatopancreas than those fed the MI‐supplemented diets. In addition, mRNA expression levels of heat shock protein 70 (Hsp70), catalase (CAT) and penaeidin were significantly differentially regulated in hepatopancreas. In summary, dietary MI supplementation may have a positive effect on improving resistance to acute hypoxia stress of L. vannamei.     

Effects of temperature,salinity and body size on the physiological responses of the Iwagaki oyster Crassostrea nippona

The physiological responses of the juvenile Crassostrea nippona in terms of filtration, oxygen consumption and ammonia excretion to changes in temperature (16–32°C), salinity (15–35 psu) and body size (small, medium and large) were investigated. In this study, the values of filtration rate (FR), oxygen consumption rate (OCR) and ammonia excretion rate (AER) increased with temperature rising from 16°C to 24°C, reaching the highest values at 24°C and 28°C; with any further increase in temperature above this limit, these values decrease drastically (p < .05). The highest Q₁₀ coefficients were 2.75 for large, 3.54 for medium at 16–20 and 3.47 for small size at 20–24°C respectively. Moreover, the responses of FR and OCR were found to be influenced significantly by salinity, tending to increase concomitantly with salinity up to 25–30 psu, though the values of these parameters were diminished dramatically (p < .05) above this level, showing a reverse pattern from that observed in AER, which firstly decreased to the lowest level at 25 and 30 psu, and then severely (p < .05) increased to the highest level at 35 psu. In addition, the low O:N ratios of all sizes of C. nippona at 16°C and 30–35 psu were indicative of a protein‐dominated catabolism, whereas the O:N ratios of large size at 20–32°C and all sizes at 20–30 psu, indicating that the metabolic energy from protein diminished and lipid and carbohydrate were used as the energy substrates. Physiological rates of C. nippona were well correlated with its size. The average values of mass exponents (b‐values) estimated in the present study were 0.657 for OCR and 0.776 for AER at different temperatures, and 0.647 for OCR and 0.767 for AER at varying salinities, signifying that physiological process of C. nippona becomes relatively slower with increasing body size regardless of temperature or salinity. Finally, our results confirm that the optimal temperature and salinity for juvenile C. nippona lie within 24–28°C and 25–30 psu respectively. The results of physiological traits in response to environmental factors of this species are informative in site selection for the cultivation.     

Characterization of a γ‐aminobutyrate type A receptor‐associated protein gene,which is involved in the response of Portunus trituberculatus to CO2‐induced ocean acidification

The γ‐aminobutyrate type A receptor‐associated protein (GABARAP) is a ubiquitin‐like modifier implicated in membrane trafficking and fusion events involving the γ‐aminobutyrate type A receptor, autophagy and apoptosis. In this study, the gene encoding GABARAP was cloned from swimming crab Portunus trituberculatus (PtGABARAP) based on the expression sequence tag (EST). The full‐length cDNA of 664 bp includes a 5′ untranslated region (UTR) of 87 bp, a 3′ UTR of 223 bp with a poly(A) tail, and an open reading frame (ORF) of 354 bp encoding a polypeptide of 117 amino acids with a predicted molecular weight of 13.96 kDa. The deduced amino acid sequence shares high similarity (93%–100%) with GABARAPs from other species and includes a conserved Atg8 domain. In a phylogenetic analysis PtGABARAP clustered with GABARAPs from other species, and more widely with other GABARAP family proteins. The impact of elevated ocean acidification (OA) on P. trituberculatus behaviours was investigated, and real‐time RT‐PCR revealed that PtGABARAP expression was up‐regulated after OA exposure. Ocean acidification also caused crabs anxiety‐like behaviours, like the shoal average speed increase, preference for dark environment (scototaxis) and fast exploration. The results indicated that GABARAP might be involved in the interactions of GABA_A receptors and elevated‐CO₂ seawater.     

Year‐round natural spawning,early development,and the effects of temperature,salinity and prey density on captive ornate goby Istigobius ornatus (Rüppell, 1830) larval survival

Natural spawning, early development and larviculture of the ornate goby Istigobius ornatus in captivity were studied for the first time. I. ornatus spawned 46 times from 31 October 2013 to 31 October 2014. Fecundity ranged from 246 to 10,214 eggs per clutch, with an average hatching rate of 77.8% ± 9.9% (M ± SEM). Fertilized eggs (1.31–1.54 × 0.46–0.50 mm in diameter) were adhesive demersal and oval‐shaped. Embryonic development lasted 84 hr at 27.5 ± 0.5°C. Newly hatched larvae [2.12 ± 0.04 mm in total length (TL)] transformed to the juvenile stage completely when TL was 7.79 mm. Effects of different water temperatures (24, 28 and 32°C) and salinities (10, 15, 20, 25, 30, 35 and 40 g/L) on per cent survivals (%) and survival activity indices (SAIs) were tested. Survival was not significantly different under different temperatures; SAIs was significantly higher at 28°C. Larvae showed the significantly higher survival and SAIs at salinities 10–30 g/L than at 35 and 40 g/L. Effect of different prey densities on survival was significantly higher in 7 days post hatch larvae fed 20 and 30 rotifers/ml. These findings could guide future programs in captive breeding technology development and commercial production of other marine ornamental gobies.     

Effects of low salinity stress on immune response and evaluating indicators of the swimming crab Portunus trituberculatus

The purpose of the present study was to investigate the effects of salinity stress on immune responses and evaluating indicators in swimming crab Portunus trituberculatus. The crabs (150 ± 8.5 g in body weight) were exposed to different salinities as 21, 26 and 31‰ (control) for 6 days. The results showed the total haemocyte counts (THC) and prophenoloxidase (proPO) activity in the haemocytes decreased significantly in the treatment groups after 6 hr, and reached the lowest levels at 12 hr. The phenoloxidase (PO) activity in the plasma increased significantly and peaked at 12 hr, then recovered to control level after 24 hr. The phagocytic per cent of haemocyte, antibacterial and bacteriolytic activities in the plasma decreased significantly in the treatment groups, and reached the lowest level at 12 hr, then recovered to control level after 72 hr. The dopamine (DA) and 5‐hydroxytryptamine (5‐HT) contents in the plasma increased significantly and peaked at 12 hr, then recovered to control level, while the noradrenaline (NE) content in the plasma had no significant change throughout the duration of the experiment. The DA and 5‐HT receptors were significantly up‐regulated in the treatment groups. The highest value of mRNA expression of DA and 5‐HT receptors occurred at 12 hr and recovered to control level after 24 hr. In addition, the cAMP and protein kinase A (PKA) contents in the haemocytes increased significantly and peaked at 12 hr, then recovered to control level after 72 hr. The phospholipase C (PLC) and protein kinase C (PKC) contents in the haemocytes increased significantly and peaked at 12 hr, then resumed to control level after 24 hr. These results speculated that biogenic amine (DA and 5‐HT) is likely to play an important role in immune modulation via cAMP/PKA signalling pathway or PLC/PKC signalling pathway when P. trituberculatus is exposed to low salinity and these results will provide scientific data for immune evaluation.     

Cottonseed meal protein hydrolysate stimulates feed intake and appetite in Chinese mitten crab,Eriocheir sinensis

One hundred and twenty crabs (initial weight: 51.32 ± 0.06 g) were fed three experimental diets with cottonseed meal protein hydrolysate (CPH) at 0, 6 and 12 g/kg for 60 days. The results showed that CPH0.6 diet significantly improved the 1‐hr feed intake, protease activity and weight gain of crabs. In thoracic ganglia, 4E‐binding protein (4E‐BP) expression level of crab fed the diets containing CPH was significantly higher than CPH0 group. In cerebral ganglia, protein kinase B and 4E‐BP expression levels of crab fed CPH0.6 diets were significantly lower than CPH0 group. In mid‐intestine, target of rapamycin (TOR) relative expression level of crabs fed the diets containing CPH was significantly lower than CPH0 group. 4E‐BP expression level showed an opposite pattern. In thoracic ganglia, cerebral ganglia and mid‐intestine, leptin receptor expression level of crabs fed CPH0.6 diet was significantly lower than CPH0 group. Neuropeptide Y and neuropeptide Y receptor expression level of crab fed CPH0.6 diet was significantly higher than CPH0 group. In conclusion, our results indicated that dietary supplementation of 6 g/kg CPH could stimulate appetite and quicken feeding rate via the TOR signalling pathway.     

Large‐scale mortality and limited expression of heat shock proteins of aestivating sea cucumbers Apostichopus japonicus after acute salinity decrease

This study deals with the mortality and related physiological responses of aestivating sea cucumber Apostichopus japonicus to acute salinity decrease. Aestivating and active sea cucumbers were exposed to a decrease in salinity (from 30 to 20 psu) at a rate of 2.5 psu every 6 h, and then maintained at 20 psu for 96 h. The mortality of aestivating sea cucumbers was ~30%, which was significantly higher than that of active sea cucumbers (~10%). This result indicated that sea cucumbers in aestivation were more susceptible to hypo‐salinity stress. To elucidate the underlying physiological mechanisms, the osmotic pressure in coelomic fluid and the levels of hsp70 and hsp90 mRNA in aestivating and active sea cucumbers were measured. No significant difference in osmoregulation was observed between the two groups. The osmotic pressure of coelomic fluid in both groups changed with decrease in ambient salinity. There were significant differences in the time course and magnitude of hsp70 and hsp90 expression between the two groups. After exposure to decreased salinity, aestivating sea cucumbers showed a delayed up‐regulation of hsp70 and hsp90 expression compared with animals in active state, and these levels decreased rapidly to control values. The expression of hsp70 and hsp90 in aestivating sea cucumbers were significantly lower than those in active sea cucumbers after salinity change. The differences in hsp70 and hsp90 expression between the states may partly explain the higher mortality of sea cucumbers in aestivation when exposed to low salinity.     

Dietary nucleotide‐rich yeast supplementation improves growth,innate immunity and intestinal morphology of Pacific white shrimp (Litopenaeus vannamei)

An 8‐week feeding trial was conducted to evaluate the effects of dietary nucleotide (NT)‐rich yeast supplementation on growth, innate immunity and intestinal morphology in Pacific white shrimp (Litopenaeus vannamei). Four isonitrogenous and isolipidic practical diets were formulated to contain 0 (control), 10, 30 and 50 g/kg of NT‐rich yeast, respectively. A total of 480 shrimp with an average initial body weight of 1.86 ± 0.02 g were randomly allocated into four groups, with four replicates per group and 30 shrimp each replicate. The results indicated that shrimp fed the diet containing 50 g/kg NT‐rich yeast had significantly higher weight gain (WG), specific growth rate (SGR) and protein efficiency ratio (PER) than those fed the control diet, and the lowest feed conversion ratio (FCR) was observed in the shrimp fed the 50 g/kg NT‐rich yeast supplemental diet. However, there was no significant difference in survival among all treatments. The crude protein of whole shrimp in the 50 g/kg NT‐rich yeast group was higher than that in the control group. Total protein, triglyceride concentrations, the activities of aspartate aminotransferase and alanine aminotransferase in serum were significantly influenced by the dietary NT‐rich yeast supplementation. The activities of serum phenoloxidase (PO) and lysozyme (LZM) of shrimp fed the diet containing 50 g/kg NT‐rich yeast were higher than those in shrimp fed the other diets. Relative expressions of alp and lzm significantly upregulated in the 30 g/kg NT‐rich yeast group compared to the control group. The intestinal fold height and fold width in the 30 g/kg NT‐rich yeast group were significantly higher than those fed the control diet; and the highest microvillus height occurred in the shrimp fed the 50 g/kg NT‐rich yeast diet. In summary, dietary 30–50 g/kg NT‐rich yeast supplementation promotes growth performance, enhances innate immunity and improves intestinal morphology of Litopenaeus vannamei.     

In vitro oocyte maturation and blood‐circulating steroid hormones in Sterlet (Acipenser ruthenus) and Siberian sturgeon (Acipenser baeri)

The oocyte of Sterlet (Acipenser ruthenus) and Siberian sturgeon (Acipenser baeri) was evaluated after incubation with 17α, 20β‐Dihydroxyprogesterone (DHP) in artificial media. Concentration of testosterone (T), progesterone (P4) and 17‐β estradiol (E2) in blood and their relation to fertilization rate were measured during experimental period. The oocytes at the same developmental stages were incubated in SIS (based on sturgeon blood ionic composition), RM2 (Ringer solution modified for sturgeons) and L‐15 (Leibovitz medium) artificial mediums in the presence of 1 μg/ml of 17α, 20β‐Dihydroxyprogesterone for 12, 18 and 24 hr. The result of this study demonstrated that fertilization rate decreased with the increasing duration of incubation in all mediums. In the Siberian and Sterlet sturgeon, the highest fertilization rate was observed in the oocytes incubated in RM2 (0.81 ± 0.4) and SIS (0.44 ± 0.8) mediums for 12 hr respectively. In the Siberian sturgeon, fertilization rate decreased significantly (p < .05) with incubation time in all three mediums. In Starlet, fertilization rate decreased significantly (p < .05) with time and reached a minimum after 24 hr (0.16 ± 0.1) in L‐15 medium. No significant (p > .05) differences were observed between blood plasma hormones and fertilization rate. The results of this study indicated that 17α, 20β‐Dihydroxyprogesterone is essential for induction of in vitro oocyte maturation in these species.     

Effects of different salinities on amino acid profile in Artemia franciscana

The effects of different salinity levels, including 10–15, 30–35, 70–75, 110–115 and 150–155 ppt, were evaluated on survival and the body amino acids (BAA) of Artemia franciscana. The results were expressed as total essential (TE), non‐essential (NE) and total amino acids (TAA); also, the ratio of the TE to NE (ENAA) on days 3 and 13 of the culture is reported. The study of changes in Artemia BAA showed that with the increase in the water salinity, the TE, NE and TAA increased significantly on days 3 and 13 of the culture. However, the highest ratio of ENAA was observed in the 110 gL^?1 salinity treatment on day 3 (1.067 ± 1.25). Regarding the effects of different salinity treatments on the survival rate of Artemia, it was observed that, generally, an increase in the water salinity would reduce the survival rate of this species. This reduction was observed on day 3 of culture in the salinity treatments above 120 (66.66 ± 1.68%) and below 30 gL^?1 (89.66 ± 0.34%) and on day 13 of culture in the salinity treatments below 30 (11.86 ± 0.13%) and above 70 gL^?1. Basically, it can be concluded that A. franciscana culture at a salinity of 110 gL^?1 can lead to the biological enrichment of Artemia in terms of amino acid profile.     

Compensatory growth,plasma hormones and metabolites in juvenile Siberian sturgeon (Acipenser baerii,Brandt 1869) subjected to fasting and re‐feeding

A 7‐week study was conducted to investigate the effect of different fasting and re‐feeding regimes on compensatory growth and some physiological parameters of juvenile Siberian sturgeon (Acipenser baerii). Fish (46.5 ± 0.5 g) were fed on a diet (containing 450 g/kg crude protein and 20 MJ/kg digestible energy) according to four feeding regimes in triplicate including: control group (C, fed everyday), W1 (2 weeks of feeding followed by 1 week of fasting and 4 weeks of re‐feeding), W2 (1 week of feeding followed by 2 weeks of fasting and 4 weeks of re‐feeding) and W3 (3 weeks of fasting followed by 4 weeks of re‐feeding). The fasted groups including W1 (119.6 ± 2.1 g), W2 (118.0 ± 1.7 g) and W3 (108.5 ± 4.8) significantly lost their weights during fasting phase and did not attain the final weight of the C (137.3 ± 1.7 g) after re‐feeding phase. The re‐feeding phase increased the specific growth rate in the fasted groups compared to the C (p < .05). After the fasting phase, concentrations of T₃, T₄, glucose, total protein and triglyceride in plasma of fasted groups were decreased, but levels of total cholesterol, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase increased compared to the C. After re‐feeding phase, except for glucose level, all mentioned metabolites were restored in the plasma of W1 group, but total protein level and aspartate aminotransferase concentrations in plasma were not restored in W2 and W3 groups. Overall, our finding demonstrated 4 weeks of re‐feeding was too short to induce full compensatory growth in A. baerii juveniles.     

The effects of salinity on reproductive development and egg and larvae survival in the spotted scat Scatophagus argus under controlled conditions

In the present study, we report the first successful instance of controlled reproduction in Scatophagus argus, which has recently emerged as a new aquaculture resource. The controlled reproduction process for S. argus was optimized with regard to salinity acclimation. Gonadal maturation was affected by salinity in both sexes. Levels of plasma 17β‐estradiol (E₂) and 11‐ketotestosterone (11‐kT) were salinity dependent and increased significantly with the duration of acclimation. Plasma levels of gonadal steroids were higher in fish held at 25‰ salinity. The highest gonadosomatic indices (GSI), 15.1 ± 1.6 in the female and 6.4 ± 1.2 in the male, were also observed at 25‰ salinity. Nevertheless, the optimal salinity for S. argus embryonic development and larval culture was 15‰. Thus, the salinity requirement for gonadal maturation and early development are quite different. The use of advanced reproductive technologies combining salinity acclimation and stimulation of luteinizing hormone‐releasing hormone analog (LHRH‐A₂) resulted in a fertilization rate of 83.2%–91.3% and embryonic survival rates of over 90%. Embryos of S. argus at the 2‐cell, blastula, gastrula and pharyngula stages were observed. Most embryos hatched after 21.0 hr of incubation at 28.0 ± 1.0°C. The development of larvae into juveniles was completed at 40–45 days posthatch (dph). In this study, we provide information about the controlled reproduction of S. argus and identify the optimal environmental parameters for S. argus embryonic and larval culture, with the aim of developing reliable reproductive techniques for its mass production.     

Pharmacokinetics and physio‐metabolic response of single and multiple dose of fenbendazole in Labeo rohita (Hamilton, 1822) fingerlings

An experiment was carried to determine the plasma fenbendazole (FBZ) concentration and physio‐metabolic responses in juveniles of Labeo rohita (90 ± 4 g) after oral administration of single doses at 10, 20 and 50 mg, 20 mg FBZ/kg b.wt. in multiple times on 1st, 3rd and 7th day. The blood samples were collected at 0.5, 1, 2, 4, 8, 12, 24, 30, 48, 72, 96 and 120 hr, after single‐dose administration, and regularly (upto 15 day) in multiple dose. Plasma FBZ concentration was determined up to the limit of detection (LoD) of 0.09 µg/ml by HPLC. There was no parent drug detected in plasma for administration of 10 mg FBZ/kg b.wt. The drug attained the peak concentration (C_max) 1.85 and 3.09 µg/ml in plasma at 4 hr (T_max) after administration of 20 and 50mg FBZ/kg b.wt. respectively. Plasma FBZ was detectable up to 96 and 120 hr with concentration 0.09 ± 0.007 and 0.098 ± 0.006 µg/ml, respectively, after single‐dose administration of 20 and 50mg/kg b.wt. In case of multiple‐dose administration, the maximum concentration of FBZ was 1.01 ± 0.03 µg/ml on 7th day that was less than to the single dose at 50 mg/kg b.wt. However, FBZ was detected up to 11 day after multiple doses. The study revealed that the hepatic antioxidant enzymes activities like superoxide dismutase, catalase and glutathione‐S‐transferase were significantly affected by increasing FBZ in single and multiple doses. The results of the present study could reveal that single‐ or multiple‐oral administration of FBZ at 20 mg/kg b.wt. in feed as antihelminthic drug in L. rohita could be considered as the safe dose.     

Prawn (Macrobrachium rosenbergii)–plant (Hydrilla verticillata) co‐culture system improves water quality,prawn production and economic benefit through stocking density and feeding regime manage

The effects and commercial implications of aquatic plant addition, and variable prawn stocking density and supply of commercial aquafeed on water quality, prawn production and economic benefit, are investigated in a prawn (Macrobrachium rosenbergii) + plant (Hydrilla verticillata) co‐culture system. Our experimental design includes a control (PM, prawn monoculture without H. verticillata, with 30 prawns/m²) and four treatments with 15% plant cover of total pond area in each replicate. Dissolved oxygen, pH, N, P, total ammoniacal nitrogen, nitrite and nitrate in this co‐culture system were significantly lower than those of PM. Prawn survival (96.2 ± 14.9%), average final weight (68.5 ± 4.9 g) and yield (879.1 ± 102.4 kg/ha) in treatment 20W80%A (20 prawns/m², 80% of control aquafeed) were obviously greater than in PM and other treatments. Despite small males (SM) and immature females (IF) being predominant in prawn + plant cultures, more than 77.2% of prawns reached or exceeded 40 g on completion of trials, and by six months were of appropriate size for market. We report culture of M. rosenbergii with H. verticillata to be both feasible and profitable. The optimal treatment, a stocking density of 20 prawns/m² with these plants, enabled reduction of commercial aquafeed to 20% conventional culture levels. For this optimal treatment, we estimate total gross revenue, profit and internal rate of return to be US$ 6,593.3 ± 103.3 ha^?1, US$ 3,095.5 ± 42.6 ha^?1 and 127.5 ± 20.7% for 20W80%A respectively; we estimate U.S. Dollar (US$) invested generates 3.87 times conventional culture revenue. Co‐culture of M. rosenbergii and plants renders prawn production ecologically and economically feasible on larger farms.     

Time‐series response of immune enzymes and catecholamines in juvenile sea cucumber Apostichopus japonicus during long‐term live transport

Sea cucumbers are free of protective exoskeleton and capable of autolysis under stress conditions. The live transport of small‐size juveniles to grown‐out sites induces hypoxia and congesting stress. The transport process is essential to production of outdoor healthy seedlings. Here, we evaluated the effects of damp transport and water transport on Apostichopus japonicus, an economic species inhabiting along the Asian coast. The survival after transport and immune performance of small‐size juveniles (0.05–0.1 g of immersed weight) were monitored during distinct duration of transport (1, 3, 5, 8 and 12 hr). The results revealed that there were no significant differences between the two transport systems within 5 hr. Given the survival for recovery of 48 hr and responses of immune enzymes and catecholamines during transport, water transport was less detrimental to sea cucumbers than damp transport when the transport duration was less than 8 hr. Once the duration prolonged to 12 hr, these immune parameters changed irregularly and the survival declined drastically. Water transport with a notably higher survival for juveniles was the more appropriate manner for long‐time transport. These findings provide insights into long‐term transport of small‐size A.  japonicus juveniles and might be beneficial for production of ecologically healthy seedlings.     

Dietary potassium requirement of fingerling Indian major carp,Labeo rohita (Hamilton), based on growth parameters,gill Na+‐K+ ATPase activity,tissue mineralization and antioxidant activities

To quantify the dietary potassium requirement of fingerling Labeo rohita (6.2 ± 0.12 cm; 1.98 ± 0.06 g), seven purified experimental diets (350 g/kg crude protein and 16.72 kJ/g gross energy) with graded levels of potassium (0.32, 1.35, 2.41, 3.46, 6.48, 9.47 and 12.39 g/kg diet) were fed to triplicate groups of fishes at 08:00, 12:00 and 16:00 hr to apparent satiation for 8 weeks. Live weight gain (LWG; 671.46%), specific growth rate (3.65%/day), protein efficiency ratio (2.16), protein gain (PG; 2.41 g/fish) and feed conversion ratio (1.32) were found to be best in fish fed diet containing 3.46 g/kg potassium. Gill Na⁺‐K⁺ ATPase activity was also highest in fish fed diet with 3.46 g/kg potassium. Potassium content of whole‐body, vertebrae and scales increased significantly with the increase in dietary potassium level up to 6.48 g/kg. Significant changes were also noted in serum malondialdehyde content, superoxide dismutase, catalase, glutathione peroxidase and alkaline phosphatase activity. Based on the maximum live weight and protein gain observed in the present study, the inclusion of 3.55 g/kg potassium is recommended for developing potassium‐balanced commercial feeds for intensive culture of fingerling L. rohita.