Effect of lactic acid on enrofloxacin pharmacokinetics in Eriocheir sinensis (Chinese mitten crab)

As a representative acidifier, lactic acid (LA) is widely used in the diets of aquatic animals. LA is the main supporter of energy metabolism and may be associated with drug metabolism. This study was designed to explore the pharmacokinetics of enrofloxacin (ENR) in Eriocheir sinensis (Chinese mitten crab) fed diets containing 0.1%, 0.2% and 0.3% LA (designated groups LA1, LA2 and LA3 respectively). The concentrations of ENR in the haemolymph, hepatopancreas and muscle were determined by HPLC after a single oral dose of 10 mg/kg. Our results showed that LA had a significant effect on the peak ENR concentrations in all the tissues (p < 0.05) by one‐way ANOVA analysis. There was a trend that C_max (peak concentration) of ENR was elevated with LA levels increasing up to 0.3% in haemolymph, hepatopancreas, muscle and T_max (time‐point of the peak concentration of the drug), t_1/2β (elimination half‐life) and AUC_(0‐∞) of ENR were shortened. Taken together, 0.3% LA might be effective in improving ENR pharmacokinetics in E. sinensis. Furthermore, it can be speculated that the enhanced biotransformation of ENR in the hepatopancreas mediated by LA is responsible for the differences in the pharmacokinetics of ENR in E. sinensis.     

The extracts of Angelica sinensis restore the digestive and absorptive capacity through improving antioxidant status in digestive organs of fish treated with trichlorfon

Firstly, a linoleic acid emulsion and fish hepatopancreas homogenate were incubated with ethoxyquin and the extracts of Angelica sinensis. The results demonstrated that ethoxyquin showed the strongest protective effects against lipid oxidation of all of the examined compounds (p < 0.05). However, ethyl acetate extract of Angelica sinensis at high concentrations showed a stronger effect on lipid oxidation than that of ethoxyquin (p < 0.05). Next, seven experimental diets that contained 0.0, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 g/kg of ethyl acetate extract of Angelica sinensis were fed to seven groups of carp (Cyprinus carpio var. Jian) respectively. After 60 days, carp were exposed to 2.4 mg trichlorfon/L in water for 4 days. The results displayed that trichlorfon exposure increased the contents of malonaldehyde and protein carbonyl in digestive organs and the activities of glutamate‐oxaloacetate and glutamate‐pyruvate transaminase in plasma, and decreased feed intake, the level of reduced glutathione, and the activities of trypsin, chymotrypsin, lipase, alpha‐amylase, Na⁺,K⁺‐ATPase, alkaline phosphatase, antisuperoxide anion, antihydroxyl radical, superoxide dismutase, glutathione reductase, and glutathione S‐transferase in digestive organs of carp (p < 0.05). Moreover, the dietary ethyl acetate extract of Angelica sinensis prevented the decrease in the above parameters in carp treated with trichlorfon (p < 0.05). These results revealed that the dietary ethyl acetate extract of Angelica sinensis could quench the trichlorfon‐induced structural and functional damage by improving the antioxidative capacity of the digestive organs of fish. Therefore, the extract of Angelica sinensis could be used as an inhibitor of trichlorfon stress in fish.     

Effects of Vibrio harveyi on plasma clotting protein (coagulogen) of white shrimp Litopenaeus vannamei

Blood clotting exhibits various important functions, including the prevention of body fluid loss and invasion of pathogens in shrimp. The effects of pathogenic Vibrio harveyi on plasma of white shrimp (Litopenaeus vannamei) in vitro and in vivo were investigated in this study. The clotting protein (coagulogen) in plasma of white shrimp pre‐incubated with extracellular products (ECP) of V. harveyi was found apparently decreased and fast‐migrated in crossed immunoelectrophoresis (CIE) gels. In addition, the coagulogen had been degraded to many low molecular‐weight protein bands in plasma pre‐incubated with ECP on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) gels. When pre‐challenged with bacterial cells and ECP of V. harveyi, the white shrimp began to die at about 30 and 16 h respectively. Moreover, plasma coagulogen was decreased more obvious in shrimp challenged with ECP than that with bacterial cells as visualized in CIE gels, and total plasma protein in both group of shrimp were all decreased. Haemolymph withdrawn from moribund shrimp pre‐challenged with V. harveyi or its ECP was observed unclottable. However, the addition of clotting factors (transglutaminase and/or Ca²⁺) to these unclottable plasma could apparently promote their re‐clotting ability as jelly‐like solid observed in microtubes. The recovery of clotting ability of plasma from moribund shrimp was due to the reformation of coagulogen (200 kDa) after adding the two clotting factors as shown on CIE and SDS‐PAGE gels. The present results suggest that the infection of V. harveyi in white shrimp may not only degrade coagulogen but also influence the presence of transglutaminase and Ca²⁺ ion.     

Examination of a practical method for copper enrichment of euryhaline rotifers (Brachionus plicatilis) as diet of Eriocheir sinensis zoea larvae

In this study, an effective method to enrich the rotifer Brachionus plicatilis with copper was developed as a feed for the Chinese mitten crab Eriocheir sinensis zoea larvae. Changes in the concentrations of other minerals in rotifers were also examined when copper was added for rotifer enrichment. The ability of Chlorella to absorb waterborne copper is much higher than that of rotifers, and hence, copper was preaccumulated in Chlorella before its ingestion by rotifers. The copper content in rotifers was comparable to the dietary copper requirement of the crab larvae when the rotifers were enriched with 0.1 mg Cu g⁻¹ Chlorella for 12 h. Further enrichment in rotifers with Cu‐enriched Chlorella and lipid emulsions did not significantly change the profile of major fatty acids and mineral composition in the rotifers. Evidence shows the feasibility of copper enrichment in rotifers using microalgae that can accumulate copper. This study indicates that copper in rotifers can be enriched by feeding copper‐enriched algae at a concentration of 0.1–0.2 mg Cu g⁻¹ Chlorella. The developmental rates of E. sinensis can be improved by feeding zoea larvae with copper‐enriched rotifers, but survival rates were not affected by dietary copper enrichment.     

Acrosome reaction of Chinese mitten-handed crab Eriocheir sinensis (Crustacea: Decapoda) spermatozoa: Promoted by long-term cryopreservation

The effects of three media, two temperatures, and fourteen durations of cryopreservation from 0 h to 450 d on in vitro acrosome reaction (AR) of spermatozoa in Chinese mitten-handed crab Eriocheir sinensis (Crustacea: Decapoda) were investigated. The spermatozoa of good quality were obtained from spermatophores by a glass homogenizer in an ice-bath and centrifugation at 4 °C. At 0 h, 2 h, 1 d, 3 d, 15 d, 30 d, 60 d, 90 d, 120 d, 150 d, 180 d, 270 d, 360 d, and 450 d of cryo-storage in Ca²⁺-free artificial seawater, 10% (v/v) glycerol, and 5% (v/v) dimethylsulfoxide at − 80 °C and in liquid nitrogen (− 196 °C), the changes in spermatozoal morphology, the time of beginning AR, and the time of maximum percentage of AR were observed. The relationships of the changes on AR presented with the different media, temperatures, and durations of cryopreservation were speculated. In this study, the cryopreserved spermatozoa all underwent AR in less than 1 h of settlement under room temperature while the percentage of AR in the control was only about 4.9%. Meanwhile, cryopreservation shortened both the time of beginning AR (from 30.11 min of the uncryopreserved spermatozoa to 0 min of cryopreserved spermatozoa on the 30th day) and the time of maximum percentage (from 59.88 min of the uncryopreserved spermatozoa to 0 min of cryopreserved spermatozoa on the 60th day). Whereas the effect of media on sperm cell AR was negligible (P > 0.05), the treatments of spermatozoa with short- and long-term cryopreservation resulted in extremely significant differences in the time of beginning AR as well as in the time of maximum percentage of AR (P < 0.01). The present data indicate that cryopreservation for long or short periods can promote the AR of sperm cells in E. sinensis and physiologically affect the ability to capacitate. It may be that the mechanism of AR in this study is the direct promotion of membrane fusion of the acrosomal cap, or destruction of the proteins inhibiting AR and activation of the proteins promoting AR, by cryopreservation. In addition, the results also show that cryopreservation can protect the spermatozoa because AR can occur in almost all sperm cells cryopreserved for less than 15 d.     

Identification of potential general markers of disease resistance in Exopalaemon carinicauda via three‐round challenge selection with an acute hepatopancreatic necrosis disease‐causing strain of Vibrio parahaemolyticus

Sixteen candidate disease‐resistant parameters were selected through which to evaluate the acute hepatopancreatic necrosis disease (AHPND)‐resistant capability of Exopalaemon carinicauda after three generations of selection for AHPND‐causing strain of Vibrio parahaemolyticus (VP_AHPND) resistance in our previous study. However, these parameters required further verification. In this study, another AHPND‐resistant E. carinicauda series was obtained through a short‐term selection procedure, consisting of three virulent challenge rounds of selection (about three‐week interval for each challenge) with VP_AHPND infection. After this selection, the survival rate at 144 hr post infection (hpi) increased from 23.33% to 37.78% and the observed 48‐hr LD₅₀ of VP_AHPND to shrimp increased from 10^5.5 cfu/ml to 10^6.5 cfu/ml. Then, the immune response of this AHPND‐resistant E. carinicauda was studied using the 16 candidate AHPND‐resistant parameters selected for in our previous study. The improved VP_AHPND clearance rate in hpi, increased total haemocyte counts, haemocyanin concentration, alkaline phosphatase activity and expressions of six immune‐related genes (Tollip and ALF in haemocytes and hepatopancreas; lysozyme, crustin and cathepsin B in hepatopancreas; and LGBP in haemocytes) at 24 hpi after the three‐round challenge selection suggest that these immune parameters may be reliable markers for the evaluation of the physiological status and potential AHPND‐resistant phenotypes in E. carinicauda.     

Effects of replacing dietary fish oil and squid liver oil with vegetable oils on the growth,tissue fatty acid profile and total carotenoids of the giant freshwater prawn, Macrobrachium rosenbergii

A feeding trial was conducted to investigate the complete substitution of either fish oil (FO) or squid liver oil (SLO) with crude palm oil (CPO), canola oil (CO) sunflower oil (SFO) or linseed oil (LO), as the sole added lipid source in diets fed to triplicate groups of giant freshwater prawn, Macrobrachium rosenbergii (initial weight = 0.42 ± 0.01 g) for 6 weeks. Prawns fed the CO or SLO diets showed significantly higher (P < 0.05) specific growth rate than those fed the FO or CPO diets. The feed conversion ratio of the prawns was significantly better when fed the CO diet, compared with the FO, CPO, SFO and LO diets. The muscle eicosapentaenoic acid content of prawns fed the vegetable oil (VO) diets were not significantly different (P > 0.05) from those fed the FO diet, although all VO‐based diets led to a significantly lower docosahexaenoic acid content compared with prawns fed the FO or SLO diet. The whole‐body total carotenoid content was significantly lower for prawns fed the SLO diet compared with prawns on the CO or CPO diets. The successful use of VO instead of marine‐based oils in prawn diets will likely reduce feeding costs associated with M. rosenbergii aquaculture.     

Impact of polyculture ecosystems of Ctenopharyngodon idellus with Eriocheir sinensis on bacterial communities in the sediments of Dongping Lake in China: Focus on optimization of polyculture modes

This study investigated the influence of polyculture ecosystems of grass carp (Ctenopharyngodon idellus) and Chinese mitten crab (Eriocheir sinensis) on the bacterial community of sediments in the Lake Dongping in China, using the Illumina HiSeq Sequencing approach. The bacterial community diversity was reduced to different extent due to the polyculture of animals. The bacterial community composition changed significantly among the ecosystems of control (without any animals cultured), G1C1 (with the highest animal biomass: C. idellus, 2 ind/m²; E. sinensis, 1 ind/m²) and G2C2 (with the lowest biomass: C. idellus, 1 ind/m²; E. sinensis, 0.5 ind/m²). Relative abundance increase for sulphide‐oxidizing bacteria and decrease for sulphate‐reducing bacteria were detected with more quantities of fish–crab cultured in ecosystems. Variations in the concentration of sediment sulphate mainly accounted for the shifts of both the bacterial diversity and community composition. Lastly, G2C2 was recommended as the optimal polyculture ecosystem that could be used in the Lake. The results suggested negative impacts of polyculture ecosystems of C. idellus with E. sinensis on the bacterial diversity but positive impacts on the community composition and indicated the role of total biomass of animas in shaping the bacterial community in the sediments of lake ecosystems.     

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.     

Development of In situ hybridization and real‐time PCR assays for the detection of Hepatospora eriocheir,a microsporidian pathogen in the Chinese mitten crab Eriocheir sinensis

A microsporidian parasite, Hepatospora eriocheir, is an emerging pathogen for the Chinese mitten crab Eriocheir sinensis. Currently, there is scant information about the way it transmits infection in the crustacean of commercial importance, including its pathogenesis, propagation and infection route in vivo. In this study, chromogenic in situ hybridization (ISH) and quantitative real‐time PCR (qPCR) assays were developed to address this pressing need, and we provided an advance in the detection methods available. Pathogens can be seen in situ with associated lesions using ISH. Positive hybridization signals were noted inside the epithelial cells of the hepatopancreas, and putative free parasite spores were observed within the tubule lumen, which were associated with lesions detected by electron microscopy and haematoxylin and eosin (H&E) analysis. qPCR allows the determination of parasite loads in infected tissues, which is important for understanding disease progression and transmission. The hepatopancreas displayed the biggest statistical copy numbers among different tissues of infected crabs, confirming a tissue‐specific pathogen infection characteristic. The qPCR assay also proved to be suitable for the diagnosis of asymptomatic carrier crabs. Combination of the two methods could facilitate the study of H. eriocheir infection mechanism in E. sinensis, enhance the early diagnosis of the pathogen and improve the management of microsporidian diseases in commercial crustaceans.     

Newly identified C‐type lysozyme in Chinese soft‐shelled turtle (Pelodiscus sinensis) exhibits potent antimicrobial activity

Lysozymes play vital roles in humoural immune response against bacterial invasion by its lytic activity. In the present study, a new C‐type lysozyme was identified and characterized from Chinese soft‐shelled turtle Pelodiscus sinensis. The full‐length cDNA of PslysC was of 923 bp, encoding a polypeptide of 148 amino acid residues. The multiple alignments and phylogenetic relationship analysis revealed the highly enzyme‐related conserved residues. The real‐time PCR analysis suggested that PslysC was constitutively expressed in a wide range of tissues with highest level in blood cells and liver. The expression of PslysC could be significantly up‐regulated under Aeromonas jandaei infection and ammonia exposure, while no significant changes were found under Poly I:C infection. The rPslysC protein was expressed in E. coli and purified by Ni‐NTA. The optimal pH and temperature for rPslysC protein lytic activities were determined at pH 7 and 30℃. rPslysC can inhibit the growth of eight kinds of Gram‐negative bacteria, and three kinds of Gram‐positive bacteria. The binding activity of rPslysC to different microbial polysaccharides and microorganism was analysed. The results showed that rPslysC could bind to selected bacteria, and exhibit a strong binding activity to lipopolysaccharide and peptidoglycan, but a weak binding activity to β‐glucan. This suggests that the binding activity might be the major mechanism of action to realize the antibacterial activity. The present study will provide helpful evidence to further understand the innate immunity of P. sinensis, and the interaction mechanisms of C‐type lysozymes with bacterial membranes.     

Role of ion channels and membrane potential in the initiation of carp sperm motility

The exposure of freshly spawned, immotile carp sperm to hypoosmotic media triggers the initiation of calcium-dependent flagellar motility. Intracellular calcium concentration has been thought to be the critical component in motility initiation, possibly acting through a novel signalling pathway. The sensitivity of sperm cells to changes of osmolality of the environment raises the question whether a mechanoregulated osmosensitive calcium pathway is involved in the activation mechanism of carp sperm motility. The sperm cells are in a depolarized state in the seminal plasma (Ψ = –2.6 ± 3 mV) and they hyperpolarize upon hypoosmosis-induced activation of motility (Ψ = –29 ± 4 mV). The intracellular sodium [Na⁺]_i, potassium [K⁺]_i and calcium [Ca²⁺]_i ion concentrations were determined in quiescent cells, and at 20, 60 and 300 s after activation. The [Na⁺]_i and [K⁺]_i of the quiescent cells were similar to the [Na⁺]_e and [K⁺]_e of the seminal plasma. Following hypoosmotic shock-induced motility, both [Na⁺]_i and [K⁺]_i decreased to one-fourth of the initial concentration. The [Ca²⁺]_i doubled at initiation of the motility of the sperm cells and remained unchanged for 5 min. Bepridil (50–250 μM), a blocker of the Na⁺/Ca²⁺ exchanger, blocked carp sperm motility reversibly. Gadolinium, a blocker of stretch-activated channels (10–20 μM), inhibited sperm motility in a dose-dependent manner and its effect was reversible. Hypoosmotic shock fluidized the membrane and gadolinium treatment made it more rigid in both quiescent cells and hypotonic shock treated but immotile sperm cells. Based on these observations, it is suggested that, besides the well-known function of potassium and calcium channels, stretch-induced conformational changes of membrane proteins are also involved in the sperm activation mechanism of common carp.     

Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gracilaria corticata in integrated culturing method under zero water exchange system

Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gracilaria corticata were measured using integrated culturing method under zero‐water exchange system in a 45‐day period. A 2 × 3 factorial design was used with two levels of shrimp stocking densities and three levels of seaweed weight densities. G. corticata was cultured on a net tied to a round polyethylene frame. Culture tanks were filled with 750‐L filtered seawater. A 40‐W compact fluorescent lamp was hung over each tank to provide adequate and sufficient light for seaweed growth. Growth parameters of shrimp and seaweed such as specific growth rate (SGR), weight gained (WG) and average daily growth (ADG) were computed based on the initial and final weight of shrimp and seaweed. The maximum and minimum SGR of L. vannamei (1.97 and 1.69%/day) were observed in treatment S₁A₃ (25 shrimp/m² and 400 g seaweed/m²) and S₂A₁ (50 shrimp/m² without seaweed) respectively. The best survival rate (94.67 ± 1.33%), WG (129.9 ± 2.9%) and feed conversion ratio (1.67 ± 0.04) were also observed in treatment S₁A₃. The SGR of G.corticata in the treatment S₁A₃ (1.97 ± 0.00%/day) was significantly higher, compared to others. Strong positive correlations were obtained between the density of G. corticata and the growth parameters of L. vannamei. The red seaweed G. corticata could boost the growth parameters, survival rate and total production of L. vannamei under zero‐water exchange system.     

Molecular cloning and characterization of the cathepsin L gene in Pelodiscus sinensis and its expression in response to bacterial challenge

Cathepsin L is one of the most important lysosomal cysteine proteases for the initiation of protein degradation, and it is involved in the immune response in many vertebrates. However, its function in the innate immune system of turtles remains poorly understood. Here, we cloned the cathepsin L gene from the Chinese soft‐shelled turtle Pelodiscus sinensis. We then examined the mRNA expression in different tissues and at different time points after infection by Aeromonas hydrophila or Vibrio parahemolyticus. The full‐length cDNA sequence cloned from P. sinensis was 1,805 bp with a 1,071 bp open reading frame, which encodes a 40.39 kDa polypeptide of 356 amino acids. The deduced protein sequence contained an active triad of Cys, His and Asn, and conserved ERWNIN and GNFD motifs. Homology analysis showed that the deduced amino acid sequence shared 77%–96% identity with other known species. Sequence alignment, phylogenetic analysis and structural comparisons revealed that cathepsin L is a member of the cathepsin family. Real‐time PCR analyses showed that turtle cathepsin L mRNA is ubiquitously expressed in various tissues, and the expression level is higher in the liver than in other tissues. The expression level in the liver peaks 24 hr after A. hydrophila infection and at two times (12 and 72 hr) after V. parahemolyticus infection. These results suggest that the cathepsin L gene of P. sinensis is likely involved in the process of the antibacterial response to A. hydrophila and V. parahemolyticus. This study is of great importance for future work exploring the molecular mechanism of antibacterial immune responses in P. sinensis.     

Pharmacokinetic/pharmacodynamic properties and P‐glycoprotein expression in pacific white shrimp,Litopenaeus vannamei after oral administration at three dosages of oxolinic acid

The experiments explored the pharmacokinetics (PK) properties of oxolinic acid (OXA) after oral administration at three dosages (10, 30 and 80 mg/kg) via medicated feed in the shrimp. The results showed that the C_max values of 4.31, 14.93 and 16.62 mg/L and AUC_0–∞ values of 92.61, 252.30 and 364.27 mg hr^?1 L^?1 were observed at three OXA dosage groups in the haemolymph respectively. In the hepatopancreas, C_max values of 7.90, 27.23 and 60.51 mg/kg and AUC_0–∞ values of 42.01, 133.06 and 219.06 mg hr^?1 L^?1 were observed at 0.5 hr post administration respectively. In the muscle, C_max values of 1.62, 5.80 and 7.36 mg/kg and the AUC_0–∞ values of 25.64, 98.10 and 134.24 mg hr^?1 L^?1 were observed at 2 hr post administration respectively. In the gills, C_max values of 2.87, 8.08 and 12.12 mg/kg and the AUC_0–∞ values of 51.38, 118.65 and 206.48 mg hr^?1 L^?1 were observed at 4 hr post administration respectively. In addition, the in vitro MIC values of OXA at three dosages against 132 strains of Vibrio were examined and showed that the minimum inhibitory concentration (MIC) values for OXA primarily ranged from 0.15–1.25 µg/ml, including eight strains of Vibrio showing MIC values ≥5 µg/ml. The MIC₅₀ and MIC₉₀ values of 132 strains were 0.62 and 1.25 μg/ml respectively. The AUC_0–24/MIC₉₀ ratios of Vibrio were 140.4 in 30 mg/kg group. Furthermore, the P‐glycoprotein (P‐gp) expression was determined in shrimp tissues after administration to three dosage groups (10, 30 and 80 mg/kg). The results showed that P‐gp expression was up‐regulated in the hepatopancreas (5.36‐, 13.68‐ and 31.06‐fold respectively) compared with the control group.     

The obvious heterosis and genetic characters of intergeneric cross and backcross juveniles between blunt snout bream (Megalobrama amblycephala) and topmouth culter (Culter alburnus)

In this study, we used artificial insemination to generate hybrid groups of fish [MC‐F₁(MA♀×CA♂) and MC‐F₂(MC‐F₁♀×♂)] by intergeneric crosses of Megalobrama amblycephala (MA) and Culter alburnus (CA); sequential backcrosses [CAM‐B₁ (CA♀×MC‐F₁♂) and MCC‐B₁ (MC‐F₁♀×CA♂)] were also performed. All these hybrids showed high rates of fertilization, hatching and survival (p > 0.05). For genetic traits, compared with those of the M. amblycephala and C. alburnus parental lines (Table 1), the fertilization rate, hatching rate and 7‐day survival rate of MC‐F1(MA♀×CA♂), MC‐F2(MC‐F1♀×♂), CAM‐B1 (CA♀×MC‐F1♂) and MCC‐B1 (MC‐F1♀×CA♂) by artificial insemination exhibited similar high rates (p > 0.05). The morphology of the four hybrids MC‐F₁/F_2, CAM‐B₁ and MCC‐B₁ were intermediate between those of their parents. Compared with their parents of MA and CA, weight gain rate (WG), specific growth rate (SGR) and protein efficiency ratio (PER) of hybrids MC‐F₁/F₂, CAM‐B₁ and MCC‐B₁ were significantly (p < 0.05) increased and feed conversion ratio (FCR) was significantly (p < 0.05) decreased after 3 months feeding. Moreover, protein content of muscle for MC‐F₁/F₂, CAM‐B₁ and MCC‐B₁ was significantly (p < 0.05) higher and carbohydrate content of muscle was significantly (p < 0.05) lower than their parents. The females and males of the four hybrids had normal gonadal development. In this study, we successfully generated intergeneric and backcross hybridization lines with fertile potential among fish of the Cultrinae subfamily and these hybrids had obvious heterosis in terms of growth performance, feed utilization and muscle quality.     

Nutritional and biochemical studies on feeding of hydrolysed and unhydrolysed detoxified Jatropha curcas protein isolate in common carp fingerlings

Detoxified Jatropha protein isolate (DJPI), prepared from Jatropha seed cake was hydrolysed using Alcalase 2.4L FG. Both DJPI and hydrolysed DJPI (HDJPI) were evaluated as a fish meal (FM) replacement in common carp (Cyprinus carpio L.) diets. The peptide concentration in HDJPI was determined using matrix‐assisted laser desorption ionization‐time of flight mass spectrometer (MALDI‐TOF). Twenty‐two peptides with molecular mass ranging from m/z 906.455 to 1774.943 Da were identified. Fingerlings (45; av. wt. 10.1 ± 0.4 g) were randomly distributed in five groups with three replicates. An 8‐week feeding experiment was conducted to compare the nutritional quality of the HDJPI, DJPI and FM. Fish were fed on iso‐nitrogenous diets: control (FM based protein), J₅₀ (50% of FM protein replaced by DJPI), J₇₅ (75% of FM protein replaced by DJPI), JH₅₀ (50% of FM protein replaced by HDJPI) and JH₇₅ (75% of FM protein replaced by HDJPI). Growth performance was higher for FM fed group compared with the other groups, while nutrient utilization parameters were similar for control and J₅₀ groups and superior (P < 0.05) to J₇₅ and both the HDJPI fed groups. No variation (P > 0.05) was observed in blood albumin and hepatosomatic index among all different groups. Blood urea nitrogen, calcium, sodium and potassium ions in the blood were similar among all five groups.