Effects of Dietary Fibre Concentrates on growth performance and digestive enzyme activities of jundiá (Rhamdia quelen)

This study was conducted to investigate the prebiotic effect of different Dietary Fibre Concentrates (DFC) (Mucilage =MG; Pectin = PN or β‐glucan + mannan = βg + M) on growth and somatic parameters, body composition and digestive enzyme activities of jundiá (Rhamdia quelen). After acclimation, fish (7.16 ± 0.06 g) were allocated into 24 tanks (30 fish per tank) and triplicate groups were fed with Control diet (0 g kg^?1 of DFC); diet supplemented with 5 g kg^?1 commercial prebiotic (CP) or diets supplemented with 5 or 10 g kg^?1 diet of MG; PN or βg + M. At the end of the trial (8 weeks), growth was significantly (P < 0.05) higher in fish fed diets supplemented with DFCs and did not differ from animals supplemented with CP. The animals that were fed Control diet presented a body protein content higher compared to those supplemented with diets containing pectin or β‐glucan + mannan (P < 0.05). However, fish fed diets added with β‐glucan + mannan yielded a higher level of protein deposited in the whole body. The activity of digestive enzymes was lower in the group supplemented with Pectin. Results indicate that supplementation with DFCs in the diet had positive effects on the performance of jundiá and are prebiotic potential candidate.     

Effect of supplementation of dietary fibre concentrates on biochemical parameters,stress response,immune response and skin mucus of jundiá (Rhamdia quelen)

The aim of this study was to assess the effects of different dietary fibre concentrates (DFC: Mucilage = MG; pectin = PN or β‐glucan+mannan = βg+M), on biochemical parameters, stress and immune response and skin mucus of jundiá (Rhamdia quelen). The fish (7.16 ± 0.06 g) were fed with Control diet (0 g/kg of DFC); diet supplemented with 5 g/kg of commercial prebiotic (CP 5) or diets supplemented with 5 or 10 g/kg of MG; PN or βg+M. After 8 weeks of the feeding trials, biochemical parameters (cholesterol, glucose, albumin and total protein), cortisol, immunoglobulin IgM and mucoproteins of skin mucus were assessed. Results demonstrated that supplementation with PN increased cholesterol levels (p<.05). After application of the stressor, most fish, except those in the PN and 10 g/kg MG groups, showed significant increases (p<.05) in cholesterol, glucose and albumin levels. The jundiás showed no difference in cortisol levels after application of the stressor (p>.05). IgM levels were significantly high in fish supplemented with DFC (p<.05). However, the concentration of mucoproteins in skin mucus was not influenced in the different treatments (p>.05). The results showed that supplementation with DFC promoted beneficial effects on the metabolism of jundiá.     

DNA barcoding reveals blend of silver catfish Rhamdia species from fish farms in Southern Brazil

Rhamdia quelen is the most produced native freshwater fish in fish farms in South Brazil. Until recently, Rhamdia branneri and Rhamdia voulezi were synonyms of R. quelen, and all the species are commercialized as silver catfish (locally called jundiá or bagre sapo) by the aquaculture industry. In fact, because these species are morphologically very similar, interspecific crosses easily might occur in fish farming. We employed standard DNA barcoding to identify jundiá molecular operational taxonomic units (MOTUs) in fish cultivated and commercialized in the industry and in possible escapees in the natural environment in southern Brazil. We analysed 48 individuals from six fish farms and 48 individuals from three rivers (Uruguay, Benedito Novo and Itapocu Rivers) using the mitochondrial cytochrome oxidase subunit I (COI). Four MOTUs were identified based on the estimated optimum threshold (OT = 0.77), and these MOTUs were concordant with Bayesian Inferece (BI) and Neighbour‐Joining (NJ) trees. Our results support the existence of at least three species in our dataset: R. branneri, R. voulezi, and R. quelen 1 and R. quelen 2. The interspecific genetic divergence ranged from 1.1% to 5.1% (mean = 3.5%), and the intraspecific distance ranged from 0% to 1.4% (mean = 0.24%). The presence of cultivated fish in the Uruguay and Benedito Novo Rivers provides evidence of genetic contamination in native populations. These results show the need to regulate aquaculture activities and to characterize the species and commercial lineages of silver catfish that are cultivated in South Brazil.     

Silver catfish Rhamdia quelen immersion anaesthesia with essential oil of Aloysia triphylla (L'Hérit) Britton or tricaine methanesulfonate: effect on stress response and antioxidant status

Responses to anaesthesia with essential oil (EO) of Aloysia triphylla (135 and 180 mg L^?1) and tricaine methanesulfonate (MS222) (150 and 300 mg L^?1) were assessed in silver catfish. Exposure to the anaesthetics elicited a stress response in the species. In the case of MS222, it was displayed as a release of cortisol into bloodstream, elevation in hematocrit and plasma ion loss. The EO presented cortisol‐blocking properties, but increased haematocrit and disturbances of hydromineral balance were observed. Liver antioxidant/oxidant status of EO and MS222‐anaesthetized silver catfish was also estimated. The synthetic anaesthetic induced lipoperoxidation, notwithstanding increased catalase contents, whereas the naturally occurring product was capable of preventing the formation of lipid peroxides, possibly due to combined actions of catalase and glutathione‐S‐transferase. Anaesthetic efficacy was also tested via induction and recovery times. Overall, the promising results obtained for the physiological parameters of the EO‐treated fish counterbalanced the slight prolonged induction time observed for 180 mg L^?1. As for 135 mg L^?1, both induction and recovery times were lengthy; despite that, the EO was able to promote oxidative protection and mitigate stress. None of the MS222 concentrations prompted such responses concomitantly.     

Effect of different dietary lipid levels on the reproduction of Rhamdia quelen (Quoy and Gaimard, 1824)

Lipids include some of the most important nutrients that affect the survival and growth of fishes in their early life stages. Lipid deficiency prior to spawning may significantly reduce egg production, hatchability and the number of surviving larvae. In this study, we investigated the effects of isocaloric diets containing 80, 140 and 200 g kg⁻¹ of lipids (lipid source, soy oil) for 90 days. The following data were collected at 0, 45 and 90 days: final weight; length; conditioning factor; hepatosomatic, gonadosomatic and visceral fat indices; total plasma protein; testosterone; 17ß‐estradiol; free amino acids; and ovary and muscle fatty acid profiles. Additionally, fecundity, oocyte production, egg morphometric parameters, and larval and postlarval growth were evaluated. Results were not statistically different for husbandry and biochemical parameters, but visceral fat content increased with increase in dietary lipid levels. The fatty acid profiles and composition differed among dietary treatments. The egg diameter and area were significantly low in fish fed the 200 g kg⁻¹ lipid diet, thereby hindering growth, survival and weight compared to those in postlarval fish (P < 0.05). The best reproductive rates were obtained when using diets containing 80–140 g kg⁻¹ total lipids.     

Effect of sex and protein level on the intermediary metabolism,growth, deposition of nutrients and profile of volatile compounds of silver catfish (Rhamdia quelen)

This study aimed at evaluating the intermediary metabolism, growth, nutritional deposition and volatile compounds of fillet of female and male silver catfish created in cages having commercial diets at two levels of crude protein. A total of 1,200 silver catfish were randomly distributed in 12 cages with initial weight of 130.05 ± 0.14 g and visually sexed. Commercial diets with 280 g/kg and 320 g/kg of crude protein were offered twice a day in the amount of 30 g/kg their biomass/day, for 90 days. At the end of the experimental period, biometry was performed to collect data, blood and tissue for later determination of plasma metabolites, hepatic metabolites, digestive enzymes, volatile compounds, deposition of nutrients and calculation of growth variables. There were metabolic differences due to the sex of silver catfish, reflecting greater productive efficiency on the females. For the plasma metabolites, in male, there were higher values of triglycerides and free amino acids than in female. For hepatic glucose, males fed with diet containing 280 g/kg of crude protein showed higher values. For protein and free amino acids, the interaction between males that received 320 g/kg of crude protein was higher. In males, there were higher values of AST and hepatic glycogen than in females. In turn, hepatic ammonia was higher in females. There was greater activity of acid protease enzymes and trypsin in the silver catfish that received diets containing 320 g/kg of crude protein. Regardless of sex, in this cultivation phase, diets with 320 g/kg of crude protein provided better performance to silver catfish. The profile of volatile compounds suggests mild odour for the fillet of silver catfish, which can be a competitive advantage of the species.     

Spawning induction in Kutum Rutilus frisii kutum (Kamenskii, 1901) using carp pituitary extract or GnRH analogue combined with metoclopramide

Kutum Rutilus frisii kutum (Kamenskii, 1901), Cyprinidae is an endemic fish of the Caspian Sea. Iranian Fisheries Organization (Shilat) produce up to 200 million fry (1–2 g body weight (b.w.)) to restock the Caspian Sea population annually. Some of these fry are produced by spawning induction in broodfish by carp pituitary extract (CPE). The objective of this study was to assay the effectiveness of the gonadotropin releasing hormone analogue (d ‐Ala⁶, Pro⁹‐Net GnRH) alone or in combination with metoclopramide (MET), a dopamine antagonist, on the percentage of ovulated females, latency period, ovulation index and fertilization success. The following hormone treatments were tested: single injection of 2 mg kg^?1 b.w. of CPE as a positive control, GnRHa alone 20 and 40 μg kg^?1 b.w. and combination of GnRHa and MET as follows: 5 μg+2.5 mg, 10 μg+ 5 mg and 20 μg+10 mg kg^?1 b.w. Negative control group was injected with 0.7% saline. The percentage of ovulated females, ovulation index and fertilization success were 90%, 71.3±1.24%, 68.4±2.3%, respectively, in the group treated with GnRHa+MET at a dose of 20 μg+10 mg kg^?1 b.w. and were significantly higher than those in the positive control (60%, 64.5±0.23%, 69.1±4.5%) (P<0.05). However, the latency period in this group was longer than that in the positive control (P<0.05). Only 20% and 40% fish ovulated in groups that received 20 or 40 μg kg^?1 b.w. GnRHa. No fish ovulated in the negative control.     

Ovulation induction in northern pike Esox lucius L. using different GnRH analogues, Ovaprim, Dagin and carp pituitary

The traditional GnRH analogue treatments applying the chemicals in pure form proved to be ineffective in inducing ovulation in northern pike (Esox lucius L). Neither mGnRHa ([D‐Ala⁶, Pro⁹NEt]‐mGnRH) nor sGnRHa ([D‐Arg⁶, Pro⁹‐Net]‐sGnRH) administered alone or together with pimozide (mGnRHa), metoclopramide (mGnRHa) or domperidone (sGnRHa) induced ovulation in females, whereas in groups receiving a carp pituitary injection most females ovulated. Spawning‐inducing agent Dagin did not induce ovulation, whereas all but one female ovulated in the carp pituitary‐treated group. Treatment with another preparation, Ovaprim, resulted in similar or lower ovulation ratio than treatment with carp pituitary. After the Ovaprim treatment, time to ovulation was not as predictable as after the carp pituitary injection. The mean fertilization rate was relatively low and similar in the groups treated with Ovaprim (54.7 ± 12.3% and 58.7 ± 19.1% for the first and second experiment respectively) and with carp pituitary (53.7 ± 10.5% and 58.9 ± 14.9%). The mean pseudogonadosomatic index (PGSI) was also similar between the Ovaprim‐treated group (14.5 ± 6.1%) and the carp pituitary‐treated one (17.9 ± 4.1%). In the present experiments, treatment with Ovaprim was less effective than that with carp pituitary.     

Retracted: Effectiveness of carp pituitary extract and luteinizing hormone releasing hormone analogue administration (by either injection or cholesterol pellet implantation) on spawning performance in female sturgeon,Huso huso

The effectiveness of common carp pituitary extract (CPE), luteinizing hormone releasing hormone analogue (LHRH‐A₂) injections and LHRH‐A₂ implants for spawning induction in female sturgeon, Huso huso was examined. In the first trial, fish were injected with 7% physiological saline (control), 50 mg kg^?1 CPE or LHRH‐A₂ at 3.5, 7, 8 or 10 μg kg^?1. In the second trial, fish were treated with LHRH‐A₂ cholesterol pellet implants containing 0, 3.5, 7, 8 and 10 μg kg^?1 LHRH‐A₂. Ovulated eggs were removed using a minimally invasive surgical technique and were artificially fertilized. Injection of CPE and LHRH‐A₂ at doses of 3.5, 7, 8 and 10 μg kg^?1 resulted in the number of ovulated fish more than LHRH‐A₂ implants (similar doses) or controls, although there was no significant difference at doses of 8 and 10 μg kg^?1 (P ≥ 0.05). The latency period of fish receiving CPE and LHRH‐A₂ injections was approximately 20 h, which was significantly lower than in fish receiving LHRH‐A₂ implants (P ≤ 0.05). Furthermore, there were no significant differences in rates of fertilization or hatching among the progeny produced in any of the treatment groups (P ≥ 0.05). In conclusion, the data from this study could be useful for artificial propagation of not‐fully‐matured females of H. huso at sturgeon hatcheries.     

Molecular characterization,inductive expression and mechanism of interleukin‐10 gene induction in the Indian major carp,catla (Catla catla)

Interleukin‐10 (IL‐10) is a multifunctional cytokine and plays an important role in diseases. In this study, IL‐10 gene was cloned and characterized from catla (Catla catla), which is a highly commercially important fish species in the Indian subcontinent. The result indicated that the full‐length catla IL‐10 (cIL‐10) gene had five exons and four introns with an open reading frame of 540 nucleotides encoding a polypeptide of 179 amino acids. A phylogenetic analysis of cIL‐10 gene sequence showed that cIL‐10 clustered with freshwater carps group as expected. Quantitative real‐time polymerase chain reaction analysis showed that cIL‐10 was expressed in gill, liver, kidney, intestine, skin and heart and its expression profile was up‐regulated in bacterial infection and LPS treatment. A close relationship of high cIL‐10 expression and low pro‐inflammatory cytokine IL‐1β expression was observed in the treated group of fish, which might reveal the role of cIL‐10 as an anti‐inflammatory cytokine. Mechanism of cIL‐10 induction was investigated by blocking nuclear factor (NF)‐κB ‐signalling with BAY 11‐7082 in catla kidney cell culture. Blocking NF‐κB suppressed IL‐10 induction by LPS, and thus it revealed that cIL‐10 was induced through NF‐κB signalling. These data could be helpful to understand the function of IL‐10 in fish in response to vaccinations, probiotics and various diseases.     

Retracted: Study of testis histology and sperm morphology in the Bunnei,Barbus sharpeyi (Gunther, 1874) induced by luteinizing hormone‐releasing hormone analogue and carp pituitary extract

This study was conducted to determine the effects of hormone treatment on testis structure in Barbus sharpeyi, as well as the morphology of sperm examined by scanning electronic microscopy (SEM). Male B. sharpeyi were divided into three groups (three fish per group) and injected with luteinizing hormone – releasing hormone analogue (LHRH–A₂) or carp pituitary extract (CPE). The first and second groups were treated with 10 μg kg^?1 LHRH–A₂ and metoclopramide (MET), and their testis were sampled pre‐ and Poststripping respectively. The third group received 2 mg kg^?1 CPE and were killed pre‐stripping. Based on the histological results obtained, the testicular connective tissue of the lumen was thicker, and seminal vesicles were of a lower volume, in fish injected with CPE in comparison to the other groups. After treatment with LHRH–A₂ and MET, not all spermatozoa within the testis were ejaculated, and only a small amount of sperm was obtained by abdominal stripping. The highest and lowest diameters of connective tissue within lobules were observed in fish receiving CPE and LHRH–A₂ treatments respectively. There was a significant difference (P < 0.05) in lobule space between the fish injected with the CPE and the fish injected with the LHRH–A₂ and MET. The SEM results revealed that the spermatozoa of B. sharpeyi were composed of a spherical to elliptical head, a cylindrical midpiece, and a lengthy flagellum. In conclusion, it was found that injection with LHRH–A₂ and MET improved the spermatogenic process in comparison to injection with CPE.     

Preliminary success using hydrogen peroxide to treat Atlantic salmon,Salmo salar L., affected with experimentally induced amoebic gill disease (AGD)

Currently, the only effective and commercially used treatment for amoebic gill disease (AGD) in farmed Tasmanian Atlantic salmon is freshwater bathing. Hydrogen peroxide (H₂O₂), commonly used throughout the aquaculture industry for a range of topical skin and gill infections, was trialled in vitro and in vivo to ascertain its potential as an alternative treatment against AGD. Under in vitro conditions, trophozoites of Neoparamoeba perurans were exposed to three concentrations of H₂O₂ in sea water (500, 1000 and 1500 mg L^?1) over four durations (10, 20, 30 and 60 min) each at two temperatures (12 and 18 °C). Trophozoite viability was assessed immediately post‐exposure and after 24 h. A concentration/duration combination of 1000 mg L^?1 for >10 min demonstrated potent amoebicidal activity. Subsequently, Atlantic salmon mildly affected with experimentally induced AGD were treated with H₂O₂ at 12 and 18 °C for 15 min at 1250 mg L^?1 and their re‐infection rate was compared to freshwater‐treated fish over 21 days. Significant differences in the percentage of filaments affected with hyperplastic lesions (in association with amoebae) and plasma osmolality were noted between treatment groups immediately post‐bath. However, the results were largely equivocal in terms of disease resolution over a 3‐week period following treatment. These data suggest that H₂O₂ treatment in sea water successfully ameliorated a clinically light case of AGD under laboratory conditions.     

Induction of ovulation in ornamental common carp (Koi, Cyprinus carpio L.) using LHRHa ([d-Ser(tBu)6, Pro9-NEt]-LHRH) combined with haloperidol and carp pituitary extract

The effects of a single injection of mammalian superactive analogue [d ‐Ser(tBu)⁶,Pro⁹‐NEt]‐LHRHa (20 μg/ kg^?1) combined with the dopamine antagonist, haloperidol (HAL, 0.5 mg kg^?1), for induction of ovulation in the koi carp broodstocks were determined under routine hatchery conditions. The results were compared with classic carp pituitary extract (CPE, double injection) application (water temperature 22 °C). Physiological saline (0.7% NaCl)‐injected fish were used as a control group and no ovulation occurred in this group. The spawning ratio was high in the LHRHa+HAL treatment group and in the CPE treatment group (6/7 and 5/7 respectively). The latency period was 14–16 h in the LHRHa+HAL treatment group and 12–14 h in the CPE treatment group (after the second injection). There was no difference between the two ovulating groups with respect to the spawning index (the weight of eggs as a percentage of female body weight) and fertilization rate of eggs (P>0.05). As a result, ovulation can be induced successfully in koi carp broodstocks with 20 μg kg^?1 [d ‐Ser(tBu)⁶,Pro⁹‐NEt]‐LHRHa+0.5 mg kg^?1 HAL treatment in a single injection without decreasing the egg quality. Application of this combination can be useful for hatchery and broodstock management in koi carp culture.     

Synchronization of ovulation in cultured northern whitefish (Coregonus peled,Gmelin 1788) using [D‐Arg6Pro9Net]‐sGnRH analogue and its effect on egg quality

Female northern whitefish were treated with salmon [D‐Arg⁶Pro⁹Net]‐sGnRHa emulsified in Freund′s incomplete adjuvant as a sustained release treatment (sGnRHa‐FIA) or with [D‐Arg⁶Pro⁹Net]‐sGnRHa dissolved in physiological saline as acute release treatment (sGnRHa‐PS). Females in four experimental groups (A, B, C, D) and one control group (E) were injected intraperitoneally as follows: A and B – sGnRHa‐FIA at doses of 50 and 25 μg kg^?1, respectively; C – double injection (DI) of sGnRHa‐PS at 25 μg kg^?1 administered 3 days apart; D – single injection (SI) of sGnRHa‐PS at 25 μg kg^?1; and E – control group receiving a 1:1 mixture of FIA and physiological saline. All treatments induced and synchronized ovulation. Mean time to ovulation was significantly reduced (P < 0.05) in GnRHa‐treated groups compared with control. Hormone treatments did not affect the relative fecundity. Slight differences were found in ovarian fluid pH between group A and D (P < 0.05). Except for group C, egg size was significantly reduced (P < 0.01) in hormonally synchronized groups compared with the control. Survival to the eyed stage of eggs from females in groups A, B and C was significantly lower (P < 0.01) than in groups D and E. Per cent hatched alevins was lower (P < 0.01) in groups A, B and C than in groups D and E. We conclude that synchronization of ovulation in northern whitefish can be induced by a single acute injection of [D‐Arg⁶Pro⁹NEt]‐sGnRHa at 25 μg kg^?1 BW if given near the natural spawning time determined by water temperature below 2°C.     

Rederivation of a mutant line (prop 1) of zebrafish Danio rerio infected with Pseudoloma neurophilia using in vitro fertilization with eggs from pathogen-free wild-type (AB) females and sperm from prop 1 males

Along with the growing number of laboratories that work with zebrafish (Danio rerio), it is necessary to have animals with good sanitary quality. Specific pathogens can interfere with the experimental results and in the life quality of the animals. Pseudoloma neurophilia is a parasite with high potential for interference in behavioural, morphology, toxicological and genetic research, and is very common in zebrafish facilities. With that, we implemented a protocol for the pathogen elimination in a genetically modified lineage (prop 1) using eggs from specific pathogen-free (SPF) wild-type fish (AB line) for in vitro fertilization, along with water recirculation equipment disinfection, appropriate PCR screening and back crossing protocols. This resulted in SPF prop 1 heterozygotes, which allowed us to move forward with subsequent crossings to develop homozygote prop 1 mutants for our research. Hence, this demonstrates a useful strategy for an individual research laboratory to rederive a specific mutant free line that is not available from other SPF laboratories.     

Detection of Herpesvirus anguillae (AngHV‐1) in European eel Anguilla anguilla (L.) originating from northern Poland—assessment of suitability of selected diagnostic methods

The Community Action Plan requests EU member states to implement measures that ensure the recovery of the severely depleted European eel stocks. One of the main threats is posed by Anguillid herpesvirus 1 (AngHV‐1) leading to increased mortality in both wild and farmed eels. Following recommendations of the OIE to minimize the risk of obtaining false‐negative results, the main aim of the study was to optimize diagnostic methods for AngHV‐1 detection using conventional PCR, nested PCR and in situ hybridization assay. While 53.3% of the individual organ samples were tested positive for AngHV‐1 by PCR, the additional virus analysis via nested PCR revealed that the actual prevalence was 93.3%. In the cell cultivation passages, a cytopathic effect was hardly found in the first two rounds. In the third passage onto cell cultures, a lytic CPE was detected. The identification and confirmation of the viruses obtained from cell cultures as well as directly from the organ tissues were proceeded by PCR, nested PCR and sequencing of the PCR products. While no positive signal was detectable in the first round by PCR using samples from the third cell culture passages, the nested PCR provided weak but visible positive signals.     

Pro‐inflammatory caspase‐1 activation during the immune response in cells from rainbow trout Oncorhynchus mykiss (Walbaum 1792) challenged with pathogen‐associated molecular patterns

In response to pathogens, the higher vertebrate innate immune system activates pro‐inflammatory caspase‐1 which is responsible for the processing and secretion of several important cytokines involved in the host's defence against infection. To date, caspase‐1 has been described in few teleost fish, and its activity has been demonstrated through substrate cleavage and inhibition by pharmacological agents. In this study, the detection of the active form of caspase‐1 during the immune response in salmonid fish is described, where two antibodies were produced. These antibodies differentially recognize the structural epitopes of the inactive pro‐caspase‐1 and the processed active form of the caspase. Firstly, caspase‐1 activation was demonstrated in vitro by ELISA, Western blotting and immunocytochemistry in rainbow trout macrophages exposed to different pathogen‐associated molecular patterns plus the pathogen Aeromonas hydrophila. This activity was clearly abrogated by a caspase inhibitor and seems to be unrelated to IL‐1β secretion. Caspase‐1 activation was then validated in vivo in gill cells from fish challenged with Aeromonas salmonicida. These results represent the first demonstration of caspase‐1 activation in salmonids, and the first evidence of the putative regulatory role which this protease plays in inflammatory response in this fish group, as described for some other teleosts and mammals.     

Characterization of β‐catenin 1 during the gonad development in the common carp (Cyprinus carpio)

β‐catenin gene is a pivotal gene for gonad development and maintenance of ovarian function in mammals. However, little is known about its expression and function in gonad development of fish. In this study, a complete cDNA (3342 bp) sequence of β‐catenin 1 was cloned from the common carp, Cyprinus carpio, by RACE PCR, which encodes a 780‐amino‐acid protein. Quantitative real‐time PCR demonstrated that β‐catenin 1 mRNA expressions were high in the testis and ovary tissue and the expression increased as the testes developed and the early stage ovaries developed. Western blot results revealed a single immunoreactive band with an estimated molecular weight of 90 kDa in testes. Immunohistochemistry analysis revealed that the β‐catenin 1 protein was concentrated mainly in the cytoplasm of early development stage of oocyte cells and in the cytomembrane of developing and mature sperm cells. 17β‐Ethinylestradiol injecting intraperitoneally into the fish decreased the relative β‐catenin 1 mRNA expression level except 1 μg/g 72 hr and 5 μg/g 48 hr of treatments in the ovary by real‐time PCR. These results suggest, for the first time, that β‐catenin 1 is an essential protein in gonad development and might be involved in ovarian early development of C. carpio.     

Dietary α‐linolenic acid affects lipid metabolism and tissue fatty acid profile and induces apoptosis in intraperitoneal adipose tissue of juvenile grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was conducted to elucidate the effects and mechanism action of dietary α‐linolenic acid (ALA, 18:3n‐3) on lipid accumulation and fatty acid profile of muscle, hepatopancreas and intraperitoneal fat (IPF) in juvenile grass carp using three isonitrogenous and isoenergetic semi‐purified diets containing 0.0% (control group), 1.0% and 2.0% ALA, respectively. The lowest intraperitoneal fat (IPF) ratio was found in 2.0% group. In the muscle, hepatopancreas and IPF, docosahexaenoic acid (DHA, 22:6n‐3) and eicosapentaenoic acid (EPA, 20:5n‐3) contents increased with the increase in dietary ALA. In the IPF, caspase 3, caspase 8 and caspase 9 showed the highest activities in 2.0% group, while the value of Bcl‐2/Bax (B‐cell leukaemia 2/Bcl‐2‐associated X protein) reached the lowest. Meanwhile, swelling of the IPF mitochondria was observed in 2.0% group. The gene expressions of fatty acid desaturase (FAD) and fatty acid elongase (ELO) in the hepatopancreas and muscle showed significantly higher levels in the treatment groups, whereas an opposite trend was existed in the IPF. Fatty acid synthase (FAS), sterol regulatory element binding protein‐1c (SREBP‐1c) in the IPF and hepatopancreas reached the lowest in 2.0% group. Overall, dietary ALA could promote n‐3 highly unsaturated fatty acids (HUFAs) synthesis and suppress the accumulation of lipid by decreasing the expression of related genes and promoting the apoptosis in IPF.