Distinct expression of GnRH genes in the red seabream brain

This paper reports the molecular cloning of a cDNA encoding the precursor of seabream gonadotropin-releasing hormone (prepro-sbGnRH) and the localization of salmon GnRH (sGnRH) and seabream GnRH (sbGnRH) expressing neurons in the brain of the red seabream (Pagrus major). The cloned prepro-sbGnRH cDNA has a 285 bps open reading frame encoding a 23 amino acid signal peptide, a 10 amino acid sbGnRH, the cleavage site (Gly-Lys-Arg), and a 59 amino acid GnRH-associated peptide. The expression of sGnRH and sbGnRH peptides, and prepro-sGnRH and prepro-sbGnRH mRNA were studied using immunocytochemistry and non-radioactive in situ hybridization, respectively. We found cell bodies that reacted positively with both the sGnRH cRNA probe and anti-sGnRH serum, but not with the sbGnRH cRNA probe or anti-sbGnRH serum in the ganglion of the terminal nerve. Cell bodies that reacted positively with the sbGnRH cRNA probe, anti-sbGnRH serum, and anti-sGnRH serum, but negatively with the sGnRH cRNA probe were found in the preoptic area (POA). Immunocytochemistry showed that a distinct bundle of axons arises in the POA which projected to the pituitary gland. These results suggest that sbGnRH is the most relevant hypophysiotropic form of GnRH.     

Biosynthesis of estradiol-17β in the ovarian follicles of the red seabream Pagrus major during vitellogenesis

ABSTRACT: The red seabream Pagrus major is a useful experimental fish for studying the endocrine control of oogenesis in teleosts. This study investigated the steroidogenic pathway for estradiol-17β (E2) biosynthesis in the ovarian follicles of red seabream. Intact follicles were isolated during vitellogenesis and incubated in vitro with different radiolabeled steroid precursors. When 17-hydroxyprogesterone (17-P), dehydroepiandrosterone (DHEA), or androstenedione (AD) were used as precursors, both testosterone (T) and estrone (E1) were synthesized by follicles, leading to estradiol-17β (E2) production. Serum steroid levels measured by enzyme-linked immunosorbent assay showed that T, E1, and E2 were present in the circulation at levels ranging from 1 ng/mL to 2 ng/mL throughout the day during the spawning season. In vitro conversion of E1 into E2, however, was 15.8-fold greater than T conversion into E2, suggesting that E2 is synthesized mainly via E1 rather than T. The results showed that E2 was synthesized from pregnenolone via 17-hydroxypregnenolone, DHEA, AD, and E1. Thus, the study demonstrated the complete steroidogenic E2 synthesis pathway in the ovarian follicles of red seabream, and revealed that E1 is the major precursor of E2.     

Effect of storage time and cryoprotectant concentrations on the fertilization rate and hatching rate of cryopreserved sperm in red seabream (Pagrus major Temminck & Schlegel, 1843)

This study examined the effects of storage time and cryoprotectant concentrations on the post‐thaw sperm of red seabream, Pagrus major. Sperm treated with 12%, 15%, 18% and 21% DMSO were cryopreserved for 10, 30, 60 and 360 days, and fertilization and hatching rates were analysed. For all groups, there were no differences in the fertilization rates and hatching rates between sperm cryopreserved for <60 days and fresh sperm (98.8±0.8%, 96.4±1.3%). However, for sperm cryopreserved for 360 days, both fertilization rates (88.6±3.0% to 7.0±1.9%) and hatching rates (79.4±7.2% to 3.3±0.8%) decreased drastically. Furthermore, the cryoprotectant concentrations affected sperm quality significantly (P<0.05). When cryopreserved for 360 days, sperm treated with 15% DMSO obtained the best results compared with other concentrations. We suggest that 15% DMSO may be an effective cryoprotectant for long‐term sperm cryopreservation of red seabream.     

Immunological detection of translation products of type V/XI collagen α1 gene and their degradation products in red sea bream muscle

We previously cloned cDNA of type V/XI collagen α1 chain (ColVa1) gene from cultured cells derived from red sea bream embryo. We raised an antibody against the deduced C-telopeptide of ColVa1 in order to detect the translation products of this cDNA and their degradation products in red sea bream muscle. To improve its specificity, the antibody was purified from rabbit antiserum by use of an affinity column cross-linked with recombinant C-terminal peptide of ColVa1 produced by Epicurian coli. The purified antibody recognized a band corresponding to the α chain of type V/XI collagen in western blot analysis of the extract of cultured cells. The antibody also recognized two bands in acid-soluble and pepsin-solubilized collagens, indicating that the translation products of the ColVa1 gene are present in muscle and that bands correspond to α and β chains of type V/XI collagen. A band corresponding to a molecular weight of approximately 65 k was detected in the NaOH extracts of muscle, suggesting that type V/XI collagen α1 chain is restrictedly digested in red sea bream muscle.     

真鲷仔稚鱼对实验微粒饲料中卵磷脂适宜需要量的研究

制备了4种n-3HUFA含量基本相等、卵磷脂含量分别为0、2．5％、5．0％、7．5％的实验微粒饲料，探讨了卵磷脂的含量对真鲷仔稚鱼生长、存活、体内相关成分以及对外部压力的耐受性的影响。25d的养殖试验结果表明，当真鲷仔稚鱼的实验微粒饲料中卵磷脂含量不超过5．0％时，随着实验微粒饲料中卵磷脂含量的增加，真鲷仔稚鱼的生长速度和成活率也相应增加。养殖试验结束后，对仔稚鱼体内相关成分的分析结果表明，仔稚鱼体内n-3HUFA和DHA的含量随着实验微粒饲料中卵磷脂含量的增加而增加。但当卵磷脂含量从5．0%增加到7．5％时，真鲷仔稚鱼的生长速度、成活率以及体内n-3HUFA和DHA的含量却没有相应增加，因此，真鲷仔稚鱼对实验微粒饲料中卵磷腊的适宜需要量应为5．0％左右。活力测试结果表明，当真鲷仔稚鱼的实验微粒饲料中卵磷脂的含量不超过5．0％时，随着实验微粒饲料中卵磷脂含量的增加，真鲷仔稚鱼对外部压力的耐受性增强，但是当实验微粒饲料中卵磷脂的含量从5．0％增加到7．5％时，真鲷仔稚鱼对外部压力的耐受性并没有继续增强，从而进一步说明真鲷仔稚鱼对实验微粒饲料中卵磷脂的适宜需要量应为5．0％左右。     

Seasonal condition and body composition changes in sharpsnout seabream (Diplodus puntazzo) raised in captivity

Changes in biological indices (weight, length, gonadosomatic index, condition factor, hepatosomatic index, intraperitoneal fat ratio and fillet portion), and body composition were examined in sharpsnout seabream (30 to 500 g) maintained in culture for one and a half years. Growth was rapid during May–September, and slow from October to April. Gonadal development took place between August and October, and gonadosomatic index peaked in October (GSI: 1.14%). The condition factor, the hepatosomatic index and the intraperitoneal fat ratio varied (CF: 1.83–2.51, HSI: 0.80–2.47%, IPF ratio: 1.27–2.95%, minimum and maximum values, respectively), falling during the warmer months of the year (May–Oct). The fillet portion (the eviscerated fish with the head, fishbone and tail removed), represented a 55% of body mass. There were inverse correlations between relative fat and moisture contents associated with growth periods and sexual maturation (fat: 13.5–18.5%, moisture: 59.9–64.6%, minimum and maximum values, respectively), whereas proportions of minerals and protein were more stable (ash: 3.8–4.8%, proteins: 17.5–21.4%, minimum and maximum values, respectively).     

Influence of temperature on muscle fibre hyperplasia and hypertrophy in larvae of blackspot seabream,Pagellus bogaraveo

The effects of temperature on Pagellus bogaraveo muscle cellularity were investigated, by morphometry, throughout the endogenous feeding stage, testing two rearing temperatures: 14 and 18 °C. The following parameters were estimated in transversal body sections at post‐opercular and post‐anal body levels: the total cross‐sectional muscle area, the total number of fibres and the mean cross‐sectional fibre area. At hatching, no significant influence of the temperature was observed on the morphometric parameters measured in the white muscle. At mouth opening, an increase in the number of post‐opercular white fibres was promoted by the highest temperature. During embryonic development, the red muscle fibre number in the post‐anal part of the larvae increased with higher temperature, but it appears that the difference was no longer present at mouth opening. An increase in the fibre area and in the total cross‐sectional area of red muscle at the post‐anal level was promoted by a 4 °C increase in the temperature during the vitelline phase. In conclusion, the axial musculature of blackspot seabream embryos/larvae reacted differently to temperature influence according to the body location, strongly supporting the need to look at and account for different body locations when evaluating muscle cellularity in fish, namely in growth/aquaculture‐related studies.     

Effects of 11-ketotestosterone and gonadotropin-releasing hormone on follicle-stimulating hormone and luteinizing hormone gene expression in castrated and sham-operated male red seabream <Emphasis Type="Italic">Pagrus major</Emphasis>

ABSTRACT:   In order to clarify the roles of androgen and gonadotropin-releasing hormone (GnRH) on gonadotropin (GTH; luteinizing hormone [LH] and follicle stimulating hormone [FSH]) synthesis, effects of castration and implantation of GnRH analog (GnRHa) or 11-ketotestosterone (11-KT) on expression of GTH subunit, α-glycoprotein subunit (αGSU), FSHβ, and LHβ genes, during the early spermatogenic stage in male red seabream Pagrus major were examined. Male red seabream underwent castration or sham-operation and were subsequently implanted with cholesterol pellets containing GnRHa, silicone capsules filled with 11-KT, or blank capsules (control). FSHβ mRNA levels increased due to castration, and it was reversed by treatment with 11-KT. 11-ketotestosterone treatment also decreased FSHβ mRNA levels in sham-operated fish. These results suggest that 11-KT acts on the pituitary to suppress FSH synthesis in male red seabream. On the other hand, neither castration nor replacement of 11-KT in castrated fish had effects on LHβ mRNA levels, whereas 11-KT treatment had slightly but significantly decreased LHβ mRNA in sham-operated fish. αGSU mRNA levels were not changed by castration or 11-KT treatment in both sham-operated and castrated fish. Meanwhile, treatment with GnRHa significantly decreased FSHβ mRNA levels in sham-operated fish, but not in castrated fish. This suggests that GnRHa may down-regulate expression of FSHβ mRNA through the production of 11-KT in testis. LHβ and αGSU mRNA levels in sham-operated fish, but not in castrated fish, were significantly elevated by treatment with GnRHa.     

First report of Streptococcus iniae in red porgy (Pagrus pagrus, L.)

This work describes the first isolation of Streptococcus iniae in red porgy, Pagrus pagrus (L.), and the first European isolation of this pathogen in gilthead seabream, Sparus aurata (L.). In both farmed fish species, infection resulted in lethargy, anorexia, abnormal swimming, exophthalmia and sudden death, with mortality rates of over 25% in red porgy and 10% in gilthead seabream. Beta-haemolytic Gram-positive cocci, catalase negative and oxidase negative, were isolated in pure culture from internal organs. Conventional and rapid identification systems, and 16S rRNA gene partial sequencing were used to identify the causative agent of the natural disease. LD50 trials were carried out to show the virulence of this isolated strain in these species, with values of 1.7 × 104 CFU per fish in red porgy and 1.32 × 105 CFU per fish in gilthead seabream. The most prominent lesions were meningoencephalitis and multifocal infiltration of macrophage cells in the kidney and spleen.     

Measuring technical,allocative and cost efficiencies of seabass and seabream farms in Greece

Abstract

A stochastic Cobb‐Douglas production frontier is used to provide estimates of output‐oriented technical efficiency, input‐oriented technical efficiency, input allocative efficiency and cost efficiency for a sample of seabass and seabream farms in Greece. Mean output‐oriented technical efficiency is estimated at 78.5%, input‐oriented technical efficiency at 73.6%, input allocative efficiency at 79.2%, and cost efficiency at 58.2%. Considering the sources of efficiency differentials among fish farms, it is evidence from the empirical results that large farms tend to achieve higher (technical and allocative) efficiency scores; specialization in either seabass or seabream affects positively technical and cost, but not allocative, efficiency; and utilization of skilled labor seems to have a positive impact only on technical efficiency.     

Purification of gonadotropins (PmGTH I and II) from red seabream (Pagrus major) and development of a homologous radioimmunoassay for PmGTH II

Two gonadotropic glycoproteins (PmGTH I and II) were purified by ion-exchange chromatography, gel filtration and preparative SDS-PAGE, from pituitaries of red seabream, a marine teleost which has an asynchronous-type ovary and spawns almost daily during the spawning season. The glycoproteins were composed of distinct subunits and the molecular weights were estimated to be 32 and 38 kDa for PmGTH I and PmGTH II, respectively. Both PmGTH I and II were active in two homologous bioassays: in vitro oocyte maturation and/or in vitro estradiol-17 production assays. These two GTHs were distinct in electrostatic properties, molecular weight, stability and yields from pituitaries during the spawning season. These properties suggest that PmGTH I and II correspond to salmon GTH I and II, respectively.A homologous radioimmunoassay with which to measure PmGTH II was developed using a rabbit antiserum against the subunit of PmGTH II and intact PmGTH II as standards and radioactive competitors. Competition curves for red seabream plasma and pituitary extract were parallel to the standard curve, while PmGTH I had low cross-reactivity (3.1 %) with the antibody. This specific RIA system showed an in vivo LHRHa induced GTH surge in the plasma of female red seabream.     

微粒饲料中鱼油n-3HUFA的分子结构对真鲷仔稚鱼生长、存活的影响

制备3种基本组成相同、添加不同类型鱼油的微粒饲料。将3种微粒饲料分别投喂真鲷(Pagrus major)仔鱼,探讨了微粒饲料中鱼油n—3HUFA的不同分子结构对真鲷仔、稚鱼生长、存活的影响。25d的养殖试验结果表明,虽然饲料2中n-3HUFA的含量高于饲料1中n—3HUFA的含量,饲料3中n—3HUFA的含量同饲料1中n—3HUFA的含量相等,但是用饲料1投喂的真鲷仔、稚鱼的全长、成活率要优于分别用饲料2和饲料3投喂的真鲷仔、稚鱼的全长、成活率。而且,用饲料1投喂的真鲷仔、稚鱼体内n-3HUFA和DHA的含量高于用饲料2和饲料3投喂的真鲷仔、稚鱼体内n—3HUFA和DHA的含量,这说明真鲷仔、稚鱼对天然鱼油的消化、吸收要优于对乙酯化鱼油的消化、吸收。     

Characteristics of a self-assembled fibrillar gel prepared from red stingray collagen

A translucent collagen gel was formed from a transparent acidic solution of red stingray collagen by adjusting to physiological ionic strength and pH in phosphate buffer and then incubating at 25–37°C. During fibril formation from red stingray collagen, the turbidity increased when the NaCl concentration was increased at constant pH and the rate of fibril formation was accelerated by higher pH or lower NaCl concentration. The T _m of red stingray collagen fibrillar gel was estimated as 44.3 ± 3.5°C, which was higher than that of the collagen solution, 33.2°C. In addition, red stingray collagen gel maintained its shape without melting and was suitable for culture of mouse stromal cells at 37°C.     

Daily change of genetic variability in hatchery offspring of red sea bream during spawning season

ABSTRACT:   In order to assess a daily change of genetic variability during spawning season, hatched larvae of red sea bream sampled on different dates were assayed by polymorphic markers such as microsatellite DNA (msDNA) and mitochondria DNA (mtDNA) control region. Based on the microsatellite loci, the average number of alleles per locus ranged between 13.7 and 18.3. The expected heterozygosities ranged between 0.843 and 0.919. A total of 23 mtDNA haplotypes were detected via digestion of mtDNA D-loop sequences with five endonucleases: Taq  I, Alu  I, Mbo  I, Rsa  I and Hinf  I. Significant fluctuation of genetic variability during spawning season was detected by both types of DNA markers. It was suggested that the genetic variability was maintained by pooling the seed fish collected on different spawning dates in a hatchery.     

Flumequine depletion from muscle plus skin tissue of gilthead seabream (Sparus aurata L.) fed flumequine medicated feed in seawater at 18 and 24 °C

We examined flumequine depletion from muscle plus skin of gilthead seabream held in seawater at 18 and 24 °C. Seven groups of 10 fish each were sampled at intervals ranging from 24 to 168 h after in-feed administration of flumequine at 35 mg/kg/day for 5 days. Muscle plus skin tissue samples were analyzed for flumequine by high-performance liquid chromatography and fluorescence detection (HPLC-SFD). Parent flumequine concentrations declined rapidly from muscle plus skin after dosing with elimination half-lives of t_1/2=22.14 and 21.43 h at 18 and 24 °C, respectively. Withdrawal periods for the maximum residue limit (MRL) of 600 μg/kg flumequine in muscle plus skin at 95% tolerance limit were 106.08 and 75.84 h at 18 and 24 °C, respectively, after treatment.     

Cost minimization and input substitution in the production of gilthead seabream

Abstract

This paper studies cost minimization decisions by utilizing a simple bioeconomic model for gilthead seabream (Sparus aurata). This model is based on a biological growth model which accounts for the effects of temperature and ration size. Although the implicit production function derived from the growth model is shown to be quasiconcave, it is not homothetic nor quasilinear. Further, technology shows increasing returns with respect to ration size and harvesting time. Optimization decisions are analyzed in two different contexts, a restricted biological setting and an expanded economic problem. It is shown that the optimal ration size increases for the latter context, reaching values commonly accepted in current industrial practice. Even though the elasticity of substitution to the shadow input prices is positive at the optimal ration size, the latter is rather robust to changes in input prices. However, optimal costs show larger sensitivity to the labor price than to the feeding and managerial costs.     

鱼粉质量对中国对虾、真鲷、饲料转化率和蛋白质消化率的影响

IntroductionInherent to the practice of intensive aquaculture is the generation of wastes having immediate and verybroad effects on the aquatic environment.There is a growing consensus about the need to reduce waste pro-duction in aquaculture to minimize the negative impacts on the environment.Nitrogen(N) and phosphorus(P) wastes are major concerns for many aquaculture operations.In the past,formulation of diets in intensive aquaculture was often aimed at meeting requirements formaximum growt…     

Restoration of the fatty acid composition of red seabream (Pagrus auratus) using a fish oil finishing diet after grow-out on plant oil based diets

This study evaluated the potential for manipulating the fatty acid composition of juvenile red seabream, Pagrus auratus. Prior to the start of the study, three groups of fish had been reared for 3 months on a fish oil based diet or diets where the added fish oil had been replaced with either canola or soybean oil. In the present study, fish that had previously been fed either the canola or soybean oil diets were fed a fish oil based diet. Three additional treatments included fish being maintained on their original diets of fish oil, canola oil or soybean oil. Fish were fed their respective diets twice daily to apparent satiety for 32 days. Samples of fish from each treatment were collected after 0, 1, 2, 4, 8, 16 and 32 days. Composition and growth of the fish were determined at each sample point. Most treatments showed no differences in growth performance, although fish fed a fish oil diet after previously being fed a soybean oil diet showed slightly better growth. No significant differences among treatments were observed in proximate composition of the fish, although there was a significant increase in total fat and individual fatty acid (g kg^?1 live‐weight) content of the fish from all treatments over the period of the study. No significant changes in the relative fatty acid composition (% of total fatty acids) over time were observed in the three treatments where fish were maintained on their original diets. In contrast, fish that were previously fed either the canola or soybean oil diets and were then fed a fish oil diet had significant changes in both the relative (% of total fatty acids) and absolute (g kg^?1 live‐weight) fatty acid content. Key changes observed included a decrease in the relative levels of polyunsaturated fatty acids (PUFA) such as 18 : 2n ? 6 and 18 : 3n ? 3. Increases in the relative levels of the long‐chain polyunsaturated fatty acids (lcPUFA) 20 : 5n ? 3 and 22 : 6n ? 3 were also observed in both treatments. The rates of absolute (g kg^?1 live‐weight) change/accumulation of these fatty acids followed an exponential equation that differed for each fatty acid in each treatment. Examination of the retention efficiency of specific fatty acids also showed marked differences between fatty acids within treatments and also differences between treatments. Biologically important fatty acids such as 20 : 5n ? 3 and 22 : 6n ? 3 had only moderate retention efficiencies and these were unaffected by treatment. In contrast, the retention efficiencies of 18 : 2n ? 6 and 18 : 3n ? 3 suggested selective retention of these fatty acids when fed fish oil diets, but moderate catabolism when fed the plant oil diets. There were also high retention efficiencies of most saturated and monounsaturated fatty acids suggestive of active retention and/or active synthesis of these fatty acids by the fish. The results of this study, particularly the increases in lcPUFA, support the usefulness of a fish oil based finisher diet for fish raised predominantly on plant oil based diets.