Dietary supplementation with taurine improves ability to resist ammonia stress in hybrid snakehead (Channa maculatus♀ × Channa argus♂)

This study determined the effect of dietary supplementation with taurine on plasma biochemical indices, blood cell apoptosis rate, survival rate and expression of catalase (CAT), glutathione reductase (GR) and heat‐shock protein 70 (HSP70) gene in juvenile hybrid snakehead under ammonia stress. Six diets were formulated to contain 0, 3.0, 6.0, 9.0, 12.0 and 15.0 g/kg taurine. Each diet was fed to triplicate groups of fish in cylindrical tanks. After 8 weeks of feeding, 20 fish per tank were exposed to ammonia stress (total ammonia nitrogen = 200 mg/L) for 48 hr. The results showed that, after ammonia stress, plasma alkaline phosphatase (ALP), triacylglycerol (TG) levels and survival rate significantly decreased (p < 0.05) while plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN), respiratory burst, blood cell apoptosis rate, hepatic CAT, GR and HSP70 mRNA expression levels significantly increased (p < 0.05). On the other hand, dietary supplementation with taurine significantly reduced levels of BUN, respiratory burst and blood cell apoptosis rate. Supplemented groups significantly increased relative mRNA expression of hepatic CAT, GR and HSP70 as well as increased survival rate (p < 0.05). These results indicated that dietary supplementation with taurine improved ability to resist ammonia stress in hybrid snakehead.     

Influence of dietary phosphorus levels on growth,body composition,metabolic response and antioxidant capacity of juvenile snakehead (Channa argus × Channa maculata)

A study was conducted to estimate the optimum requirement of dietary phosphorus (P) for Channa argus × Channa maculata. Effects of dietary P levels on the tissue composition, serum biochemical parameters and antioxidant status were also examined. Five practical diets were formulated to contain graded levels (4.8 g kg^?1, 6.4 g kg^?1, 7.9 g kg^?1, 9.4 g kg^?1 and 11.0 g kg^?1) of available P from dietary ingredients and monocalcium phosphate (MCP). Each diet was randomly assigned to triplicate groups of 30 juvenile fish (initial body weight, 20.50 ± 0.53 g) for 8 weeks. The results showed that the specific growth rate (SGR) and weight gain (WG) were all significantly improved by dietary P up to 9.4 g kg^?1 (P < 0.05) and then levelled off beyond this level. Broken‐line analysis showed maximum weight gain (WG) was obtained at dietary available P concentrations of 9.6 g kg^?1. With the increase in dietary P level, protein efficiency rate (PER) increased significantly and reached a plateau, while the feed conversion ratio (FCR), the mesenteric lipid somatic index (MSI) and the whole‐body lipid content significantly reduced (P < 0.05). Dietary P levels also affected the mineralization (ash and P) of whole body, vertebrae and scale (P < 0.05). Quadratic analysis based on P contents in whole body, vertebrae, scale and ash content in vertebra indicated that the available P requirements were 10.4, 9.8, 10.0 and 10.3 g kg^?1, respectively. However, no differences were found in the whole‐body moisture, crude protein, serum calcium (Ca) contents or Ca/P value, as well as the viscerosomatic index (VSI) and hepatosomatic index (HSI) among all the treatments (P > 0.05). Triglyceride (TG), total cholesterol (TC), high‐density lipoprotein cholesterol (HDL‐C) and low‐density lipoprotein cholesterol (LDL‐C) decreased significantly, while serum P content, HDL‐C/TC and HDL‐C/LDL‐C value increased significantly with dietary available P levels (P < 0.05). No significant changes in superoxide dismutase activity and malondialdehyde (MDA) content were observed (P > 0.05), but serum catalase (CAT) and glutathione peroxidase (GPx) activities and the ratio of CAT/SOD and GPx/SOD increased significantly with increasing dietary P levels (P < 0.05). In conclusion, the optimal P requirement of juvenile snakehead in practical feed was 9.6 g kg^?1. Signs of P deficiency were characterized by poor growth, slightly reduced mineralization and the antioxidant capacity and an increase in body lipid content.     

Ploidy polymorphism and morphological variation among reciprocal hybrids of Pseudosciaena crocea (♀) × Miichthys miiuy (♂)

Diploid and triploid hybrids of large yellow croaker (Pseudosciaena crocea, Pc) and Miiuy croaker (Miichthys miiuy, Mm) were produced by interspecific hybridization. The result showed that Pc (♀) and Mm (♂) can be successfully crossed and hybrid offspring of normal hatch rate were produced. In contrast, a lower fertilize and hatch rate were produced for hybrids of Pc (♂) and Mm (♀), and the offspring died day 19 after hatching. To confirm the ploidy of hybrid offspring, we examined the karyotypes of offspring. The result of DNA relative content investigation and genetic relationship analysis demonstrated that the major hybrid offspring were hybrid diploid. In addition, a small number of hybrid were triploid and gynogenesis diploid. Morphological analysis based on the measurable traits data indicated the success of the interspecific crosses. The natural gynogenetic diploid large yellow croaker (PP), diploid hybrids (PM) and triploid hybrids (PPM) were closer to maternal parent (Pc) than paternal parent (Mm). This study provided genetic evidences of the chromosome, DNA content and morphological levels to support the successful establishment of the polyploidy hybrids of Pc (♀) and Mm (♂).     

Responses of hybrid sturgeon (Huso dauricus ♀ × Acipenser schrenckii ♂) to oral administration of phosphorus

To better understand the phosphorus (P) utilization in hybrid sturgeon (Huso dauricus ♀ × Acipenser schrenckii ♂), a combined technique with oesophageal intubation, dorsal aorta cannulation and urinary catheterization was used in the present study. Groups of five hybrid sturgeon were orally administrated with graded doses of P (0, 50, 100 and 200 mg P/kg BW) in the form of monocalcium phosphate. The high dose of P (200 mg P/kg BW) caused a significant increase in plasma P concentration. Plasma calcium (Ca) concentrations were significantly increased in the fish treated with 0 and 100 mg P/kg BW. Both P and urea excretion showed increased changing pattern with increasing P intubation level and exhibited significant higher value in 200 mg P/kg BW group. A significant increase in Ca excretion was found in the group without P supplementation (0 mg P/kg BW). ALP and SOD activity in plasma was inhibited in fish at P‐deficient groups (0 and 50 mg P/kg BW). The data in the current study showed that hybrid sturgeon at 100 mg P/kg BW treatment exhibited better P and Ca utilizations with lower waste excretion, as well as favourable status of anti‐oxidation.     

The optimum dietary isoleucine requirement of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)

A six‐week growth trial was conducted to evaluate the optimum dietary isoleucine requirement of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). Seven isoenergetic (3,400 kcal/kg of dry matter), isoproteic (496 g/kg of dry matter) and isolipidic (70 g/kg of dry matter) diets were formulated to contain graded Ile levels (7.3, 11.3, 15.7, 19.6, 23.5, 26.8 and 30.8 g/kg, dry‐matter basis). Each experimental diet was fed to triplicate groups of 12 hybrid grouper juveniles (average initial body weight: 6.00 ± 0.01 g/fish). Experimental fish were randomly distributed into 21 glass tanks (L 60 × W 45 × H 50 cm) connected to mechanical and biological water filters as a recycling system. Fish were fed twice daily (08:00 and 16:00) to apparent satiation. After the sampling of the growth trial, the remaining fish in each group were fed their corresponding diets for 2 d and then exposed to 4 mg Cu²⁺ · L^?1 water for 24 hr. Results showed that growth performance and feed utilization were significantly affected by different dietary Ile levels (p < .05). Weight gain percentage (WG%), protein productive value (PPV), protein efficiency ratio (PER) and feed efficiency (FE) were increased as dietary Ile level increased, reaching a peak value at 19.6 g/kg dietary Ile, and thereafter, these four parameters declined as dietary Ile level continued to increase. Daily feed intake (DFI) showed an opposite tendency of variations as FE. The quadratic regression analysis of WG%, PPV, PER and FE against dietary Ile levels indicated that the optimum dietary Ile requirement for hybrid grouper was estimated to be 19.8, 20.8, 19.4 and 19.1 g/kg dry matter, respectively. Among all experimental treatments, fish fed 19.6 g/kg dietary Ile had the highest expression of growth and protein synthesis‐related genes, including growth hormone (GH) in pituitary, insulin‐like growth factor 1 (IGF‐1), growth hormone receptor 1 (GHR1), target of rapamycin (TOR) and S6‐kinase 1 (S6K1) in liver. Gut micromorphology was significantly influenced by dietary Ile levels. After the exposure to 4 mg Cu²⁺ · L^?1 water for 24 hr, fish fed 19.6 g/kg dietary Ile had the highest survival and the best immunologic manifestation among all experimental treatments. Generally, the optimum dietary Ile requirement for maximum growth of hybrid grouper was estimated to be 19.8 g/kg dry matter, corresponding to 39.9 g/kg dietary protein.     

Dietary myo‐inositol requirements of juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenckii ♂)

This study was conducted to determine the optimal dietary myo‐inositol (MI) requirement of juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenckii ♂). Fish with an initial average body weight of 28.28 ± 0.41 g were randomly divided into six groups with three repetitions in each group and fed the experimental diets with different graded MI concentrations (70.2, 130.0, 270.2, 443.0, 1,096.6 and 1997.3 mg/kg) three times per day for 12 weeks. The results showed that with the increase in dietary MI level from 70.2 mg/kg to 443.0 mg/kg, weight gain (WG) was increased significantly (p < .05); however, when the MI level was further increased, there was no significant difference in weight gain rate (WG) over time (p > .05). Specific growth rate (SGR) showed a similar trend. The feed conversion ratio (FCR), survival rate (SR), hepatosomatic index (HSI), viscerasomatic index (VSI) and condition factor (CF) were not affected by dietary MI supplementation (p > .05). Dietary MI did not affect whole‐body and muscle crude protein and ash content (p > .05), while dietary MI significantly affected the muscle and whole‐body crude lipid and moisture content (p < .05). Moreover, serum triglyceride (TG), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were decreased by increasing dietary MI levels. Total antioxidant capacity (T‐AOC), malonic dialdehyde (MDA) and superoxide dismutase (SOD) activities in the liver were also affected by the dietary MI (p < .05). In summary, this study indicated that the optimal dietary MI requirement for juvenile hybrid sturgeon (Acipenser baerii ♀ × A. schrenckii ♂) was 335.84 mg/kg and 590.65 mg/kg based on the maximum WG and liver MI accumulation indices respectively.     

Molecular cloning,characterization, tissue expression and nutritional regulation of O‐GlcNAc transferase gene in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂)

O‐GlcNAc transferase gene (OGT) was considered as the sole rate‐limiting enzyme in the O‐GlcNAc modification. In the present study, the OGT gene of hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) was cloned and characterized, and its expression in response to dietary carbohydrate level and acute glucose treatment was investigated. The full‐length of OGT (GenBank accession no. KY656469 ) was 4,063 bp, including a 302 bp 5′untranslated terminal region (UTR), a 3,165 bp coding region that encoded 1,054 amino acids residues and a 596 bp 3′ UTR. The highly conservation of OGT gene between fish and mammals was also observed through multiple sequences alignment and phylogenetic analysis. O‐GlcNAc transferase gene was ubiquitously expressed in all detected tissues with highest expressions in brain and liver, to a lesser degree, in eye, heart, kidney and intestine. The increasing dietary carbohydrate from 8.02% to 16.08% had no significant effect on the mRNA expression of OGT. However, the expression of OGT was slightly elevated at 6 hr post‐glucose injection, and the elevation became significant at 24 hr time‐point. These data may enhance our understanding on the nutritional regulation of OGT and O‐GlcNAc modification in fish species.     

Fishmeal replacement by soybean,rapeseed and cottonseed meals in hybrid sturgeon Acipenser baerii ♀ × Acipenser schrenckii ♂

A 12‐week feeding trial was conducted to evaluate the effects of replacing fishmeal (FM) with soybean meal (SBM), rapeseed meal (RM) and cottonseed meal (CSM) on growth, feed utilization and body composition of juvenile hybrid sturgeon Acipenser baerii ♀ × Acipenser schrenckii ♂ (initial body weight, 8.63 ± 0.24 g). Five isonitrogenous and isoenergetic diets were formulated as follows: a control diet (FM60) containing 600 g/kg FM and four other diets (FM45, FM30, FM15 and FM0 containing 450, 300, 150 and 0 g/kg FM, respectively) where protein from FM was substituted by a mixture of SBM, RM and CSM. Fish fed FM0 and FM15 had poorer growth performance, feed utilization, apparent digestibility coefficients of dry matter, protein, lipid and gross energy, and fed FM0 had poorer hepatosomatic index and survival compared with the fish fed FM60. The whole body lipid in fish fed FM0 was significantly higher than that in fish fed FM60 and FM15. This study indicates that 300 g/kg of FM can be replaced with a mixture of SBM, RM and CSM in the diet of juvenile hybrid sturgeon without compromising growth performance, feed utilization and body composition.     

Effects of dietary α‐ketoglutarate supplementation on the antioxidant defense system and HSP 70 and HSP 90 gene expression of hybrid sturgeon Acipenser schrenckii ♀ × A. baerii ♂ exposed to ammonia‐N stress

This study was conducted to determine the effects of dietary α‐ketoglutarate (AKG) supplementation on the antioxidant defense system and gene expression of heat shock protein (HSP) 70 and HSP 90 in hybrid sturgeons Acipenser schrenckii ♀ × A. baerii ♂ exposed to ammonia‐N stress. A 2 × 3 factorial experiment was arranged, in which each diet (0%, 1% AKG) was randomly assigned to 0.25 (control) 5 and 10 mg L^?1 ammonia‐N groups with three replicate aquaria for each 72 h. The 10 mg L^?1 ammonia‐N significantly increased serum ammonia concentrations and intestinal Gln concentrations and GS activity compared with the 0.25 or 5 mg L^?1 ammonia‐N groups. The intestinal Gln concentration and GS activity increased, and the serum ammonia concentration decreased, in fish given dietary supplementation of 1.0% AKG compared with fish given diets without AKG. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity in serum, gills and intestines decreased when fish were exposed to 5 or 10 mg L^?1 ammonia‐N, and their activity increased in fish given diets with 1% AKG. Catalase in the serum and gills decreased when fish were exposed to 5 or 10 mg L^?1 ammonia‐N and increased in fish given diets with 1% AKG. The 10 mg L^?1 ammonia‐N or 1% AKG supplementation increased HSP 70 and HSP 90 gene expression in the liver. The increased activity of antioxidant enzymes, and increased HSP 70 and HSP 90 gene expression in fish fed diets containing 1% AKG suggested higher tolerance to ammonia‐N stress.     

草鱼胰岛素样生长因子1受体基因cDNA全序列的克隆及功能

本研究采用cDNA末端快速扩增法(RACE)克隆了草鱼(Ctenopharyngodon idellus)IGF-IR基因全长cDNA序列, 并对该基因在草鱼不同时期胚胎和成鱼不同组织中的表达进行了分析。序列分析表明, 草鱼IGF-IR基因cDNA序列全长5 741 bp, 包括5′端非翻译区822 bp, 3′端非翻译区581 bp, 开放阅读框4 338 bp, 共编码1 445个氨基酸。序列比对结果显示, 草鱼IGF-IR可能属于a型, 该基因编码的氨基酸序列与鲤(Cyprinus carpio)IGF-IRa、斑马鱼(Danio rerio)IGF-IRa和人类(Homo sapiens)IGF-IR的相似性分别为95%、93%和66%, 具有较高的同源性, 表明该基因在长期进化中具有较高的保守性。RT-PCR结果表明, 该基因从16 hpf(hours post fertilization)胚胎期到出苗期都有表达, 在成鱼大部分组织中均有表达。原位杂交结果显示, 草鱼IGF-IR mRNA在不同时期胚胎组织中广泛存在, 其中在脑部、脊索和尾部等生长旺盛组织的细胞中表达量较高。本研究为进一步探索草鱼IGF-IR基因在生长发育信号通路中的作用和育种提供了基础资料。

     

Weaning methods using formulated feeds for snakehead (Channa striata and Channa micropeltes) larvae

The culture of snakehead fish (Channa striata and Channa micropeltes) in Vietnam is limited, and snakehead culture has been banned in Cambodia, because traditional practices include capture of fingerlings from the wild as seed, as well as capture of small‐size (also known as trash fish or low‐value) fish. As hatchery breeding technology has improved, we investigated the optimal weaning practices for these two species. Both laboratory experiments and farm trials were conducted. For C. striata, the optimal weaning procedure is to begin at 17 days after hatch (dah) and wean the fish at 10% replacement of live feed with formulated feed per day. However, for C. micropeltes, the optimal procedure is to wait until 40 dah to begin weaning and then to wean the fish with a 10% replacement of live feed with formulated feed every 3 days. These results should enable farmers to domesticate snakehead culture in Vietnam and Cambodia and eliminate reliance on fish captured from the wild as both seed and feed.     

Effects of live and artificial feeds on the growth,digestion, immunity and intestinal microflora of mandarin fish hybrid (Siniperca chuatsi ♀ × Siniperca scherzeri ♂)

How to acclimate mandarin fish to eat artificial feeds has been always a challenge for researchers. The mandarin fish after hybridization (Siniperca chuatsi ♀ × Siniperca scherzeri ♂) could be fed artificial feeds which solved the problem to some extent. However, the growth performance, digestibility, immunity and intestinal microflora of mandarin fish hybrid fed artificial feeds need further study. One hundred and twenty fish of similar size (average weight, 19.5 ± 0.9 g) were randomly stocked into six 250‐L aquariums and separately fed live baits (the control) or artificial feeds in triplicate for 70 days. The weight gain and special growth rate of fish fed artificial feeds were significantly lower than those of fish fed live baits (P < 0.05). The protease activities of stomach, liver and intestine in fish fed live baits were all significantly higher than those in fish fed artificial feeds. The activities of catalase and lysozyme, the content of glutathione in serum of fish fed live baits were all significantly higher than those in fish fed artificial feeds. However, the content of malondialdehyde in liver of fish fed artificial feeds was significantly higher than that in fish fed live baits. The dominant bacteria in both groups were Proteobacteria, Bacteroidetes, Planctomycetes, Cyanobacteria, Verrucomicrobia. However, live baits greatly affected the amount of beneficial and harmful bacteria of intestine in mandarin fish hybrid and broke the balance of intestinal flora.     

The effects of water temperature and stocking density on survival,feeding and growth of the juveniles of the hybrid yellow catfish from Pelteobagrus fulvidraco (♀) × Pelteobagrus vachelli (♂)

In this study, we aimed to investigate the effects of water temperature and stocking density on the survival, feeding and growth of the juveniles of the hybrid yellow catfish from Pelteobagrus fulvidraco (♀) × Pelteobagrus vachelli (♂) using the parameters as follows: survival rate (%), feeding rate (% day^?1), feed conversion ratio, specific growth rate (% day^?1), coefficient of variation (%), productivity (P, g m^?3 day^?1) and condition factor. We reared the juvenile fish (3.25 ± 0.21 g) at 12 water temperature levels and six stocking density levels (each level included three aquaria in two batches of experiments). The results showed that all groups survived at a temperature range of ≤35°C during a 46‐day experimental period, and they could achieve a high growth at a water temperature range of 26–32°C. The optimal temperature for growth was 29.8°C. Productivity peaked at a stocking density of 1.9 kg m^?3. Our results indicated that the hybrid is very suitable for commercial aquaculture.     

Use of response surface methodology to study the combined effects of temperature and salinity on hatching and deformity of the hybrid grouper,Epinephelus fuscoguttatus (♀) × Epinephelus polyphekadion (♂)

This study seeks to identify the optimal combination of temperature and salinity for the embryo development of the hybrid groupers, Epinephelus fuscoguttatus (♀) × Epinephelus polyphekadion (♂), using the central composite design and response surface methodology. Results have shown that the low hatching rates of fertilized eggs and high deformity rates of larvae were the result of lower or higher temperature/salinity levels. The linear effects of temperature and salinity were significant in both hatching rates and deformity rates (p < .01). Similar results were observed in the quadratic effects of temperature and salinity (p < .01). Effects of temperature were positively related to salinity effects (p < .05). Salinity effects were more remarkable than temperature on hatching rates, but not the same on deformity rates. Model equations were established for temperature and salinity effects on hatching rates and deformity rates. A high coefficient of determination (R² > .97) was found for the adequacy and predictive capability of model equations, indicating that these models can be applied to prediction. The optimal combination of temperature and salinity was 27.58°C/30.94, which was derived from the statistical optimization approach. In this condition, the hatching rate was the highest at 85.73%, and the deformity rate was the lowest at 5.96%, with a high desirability function of 0.957. The results of this study can be applied to improve hatching and decrease deformity in future cultures of hybrid groupers.     

Effects of alternative dietary lipid sources on growth performance,health status and fillet fatty acid composition of hybrid sturgeon (Acipenser baeri Brandt ♀ × Acipenser schrenckii Brandt ♂)

An 8‐week trial was conducted to determine the effects of total replacement of 12.9% fish oil (FO) with soybean oil (SBO), peanut oil (PNO), sunflower seed oil (SFSO), corn oil (CO) and canola oil (CNO) on growth performance, health status and fillet fatty acid composition of hybrid sturgeon (194.28 ± 0.14 g). Compared to the FO group, dietary SBO decreased growth performance (p < .05), increased serum glucose and hepatic lipid content (p < .05). No obvious adverse effects on growth performance and health status were observed in PNO, SFSO and CO groups (p > 0.05). The fish fed with CNO had increased growth performance (p < .05), reduced serum ALT, AST, LDL‐C (p < .05) and enhanced serum GSH‐Px, T‐AOC, and LZM, MPO, C4 (p < .05). The contents of C18:1n9, C18:2n6, and ∑n‐3 PUFA and ∑n‐6 PUFA in fillets showed a positive linear correlation with the diets (p < .05). In summary, PNO, SFSO and CO are probable alternative lipid sources to fully replace FO. Hybrid sturgeon prefers to use CNO as a lipid source with improved growth performance and health status. The fillet fatty acid composition mirrors the dietary fatty acid composition.     

Evaluation of dietary vitamin E supplementation on growth performance and antioxidant status in hybrid snakehead (Channa argus × Channa maculata)

An 8‐week feeding trial was conducted to investigate the effects of dietary vitamin E on growth performance and antioxidant status of juvenile snakehead. The snakeheads (20.47 ± 0.06 g) were fed with five isonitrogenous and isoenergetic experimental diets that contained 13 (the basal diet), 52, 79, 168 and 326 mg of vitamin E kg^?1, respectively. The maximum specific growth rate (SGR) and feed intake (FI) were achieved in fish fed on a diet with 79 mg kg^?1 vitamin E (p < .05). No significant differences were found in protein efficiency ratio (PER), feed conversion ratio (FCR) and survival of fish among all groups (p > .05). Vitamin E supplementation improved hepatic glutathione peroxidase (GPx) activity significantly. A consistent decline in the hepatic and serum malondialdehyde (MDA) content was observed in fish fed diets with the increased supplementation of vitamin E (p < .05). In addition, with the increasing level of vitamin E, serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were reduced (p < .05). However, serum superoxide dismutase (SOD), catalase (CAT) and alkaline phosphatase (ALP) activities were enhanced in fish fed 79 mg vitamin E kg^?1 diet and then decreased significantly as the content of vitamin E in the diet increased (p < .05). Meanwhile, serum albumin (ALB) and globulin (GLB) were not affected by the supplemental levels of dietary vitamin E (p > .05). The vitamin E concentrations in liver and serum increased significantly with increasing dietary vitamin E (p < .05). Based on the broken‐line regression of SGR, vitamin E level in the diet is estimated to be 80.5 mg kg^?1 for Channa argus × Channa maculata. In conclusion, this study indicated that the dietary appropriate vitamin E could enhance the growth performance, antioxidant status and non‐specific immune response.