Molecular characterization and expression pattern of AFPIV during embryogenesis in gibel carp(Carassiu auratus gibelio)

As a new type of AFPs, AFPIV has been firstly identified in longhorn sculpin (Myoxocephalus octodecimspinosus), and in recent years, its cDNA and amino acid sequence have been reported, and its pancreatic synthesis has been firstly reported in polar fish. However, its expression patterns during fish embryogenesis have not been elucidated yet. By differential screening, we cloned the CagAFPIV in gibel carp, Carassius auratus gibelio, demonstrated its predominant expression during embryogenesis. RT-PCR detection revealed that CagAFPIV was first transcribed from blastula stage and kept a high level during embryogenesis and declined remarkably in hatched larva. In situ hybridization revealed that CagAFPIV transcripts were firstly distributed over the margin and marginal blastomere in blastula stage embryos, at the early-gastrula stage the positive signals distributed in the marginal cells and the internalization cells, and later restricted to the cells the yolk syncytial layer (YSL) from later gastrula stage to larva stage. Consistently, the CagAFPIV protein also kept a high level during embryogenesis, and the high protein level retained some days after the larva hatched. Our work, for the first time, revealed the dynamic expression and distribution of CagAFPIV during embryogenesis.

Effect of high dietary starch levels on the growth performance, blood chemistry and body composition of gibel carp (Carassius auratus var. gibelio)

An 8-week growth trial was carried out in a semi-recirculation system to investigate the effect of high dietary starch levels on the growth performance, blood chemistry, starch utilization and body composition of gibel carp (Carassius auratus var. gibelio). Five isonitrogenous and isocarloric experimental diets were formulated to contain different starch levels (24%, 28%, 32%, 36% and 40% respectively). Triplicate groups of fish (24 fish per tank with an average body weight, of 8.5 g) were assigned to each diet. The results showed that dietary carbohydrate levels significantly affected the growth performance, hepatopancreatic lipid content, pyruvate kinase (PK) activity and whole-body lipid content. Growth performance, body crude lipid and plasma glucose concentrations showed a decreasing trend with an increase in dietary starch from 24% to 40%. Pyruvate kinase activities and hepatopancreatic lipid content showed an increasing trend with the dietary starch increasing from 24% to 32%, and then a decreasing trend with the dietary starch increasing from 32% to 40%. No significant difference in the hepatopancreatic hexokinase (HK) activity, plasma triglyceride contents, body crude protein, ash and calcium (Ca) and phosphorus (P) contents was observed between different treatments. In conclusion, higher dietary starch levels (32-40%) significantly (P < 0.05) decreased the growth of gibel carp in the present study.

Genetic diversity in crucian carp (Carassius auratus)

A survey of restriction fragment polymorphism in mitochondrial DNA of three subspecies of Carassius auratus throughout four provinces in China was undertaken using 17 restriction enzymes. Two carp, Cyprinus carpio rubbrofuscus and Cyprinus carpio carpio, were included as the outgroup. A total of 16 haplotypes was observed: 5 in tetraploids of C. auratus auratus; 8 in hexaploids of C. auratus auratus; and 2 in C. auratus gibelio and C. auratus cuvieri, respectively. The tetraploids and hexaploids share three common haplotypes as I, V, and VI. C. a. Cuvieri may have diverged first among the three subspecies. Interestingly, C. a. auratus and C. a. cuvieri did not form monophyletic clades, which indicated that the classification of carassius auratus required further studies. The current hypothesis, that hexaploids originated from tetraploids by a polyploidy event, is less favorable, based on the distribution of haplotypes and the lower diversity in tetraploids than in hexaploids. Our data also indicate that divergence of hexaploids and tetraploids might be recent and mtDNA polymorphism existed before the divergence. Meanwhile, genetic isolation exists between the hexaploids and the tetraploids.

Positive selection on multiple antique allelic lineages of transferrin in the polyploid Carassius auratus

Transferrin polymorphism has been studied in the polyploid Carassius auratus by cloning and sequence analysis of cDNAs from its three subspecies C. auratus gibelio, C. auratus auratus, and C. auratus cuvieri. DNA polymorphism of extremely high extent was shown for the transferrin gene by the 248 segregation sites among coding region sequences of its alleles. The deduced amino acid sequences of the transferrin alleles showed variable theoretical physicochemical parameters, which might constitute molecular basis for their electrophoretic heterogeneity. Positive selection was inferred by the replacement/synonymous ratios larger than 1 in partial allelic lineages which was subsequently confirmed by likelihood simulation under neutral or selection models. Furthermore, the correspondent sites to these selected codons were collectively located at two planes in the crystallographic structure of rabbit transferrin, which suggested that the rapid evolution of C. auratus transferrin might correlate to its adaptation to variable environmental elements such as oxygen pressure. The minimal 26 recombination events were detected among coding sequences of C. auratus transferrin, with partial mosaic sequences and breakpoints identified by identity scanning and information site analyses. Phylogenetic analyses revealed multiple antique allelic lineages of transferrin, which was estimated to diverge fifteen to twenty MYA. All these features strongly suggested the role of balancing selection in long persistence of high transferrin polymorphism in C. auratus. Furthermore, owing to its particular evolutionary backgrounds, the silver crucian carp might possess a distinctive balancing selection mechanism.

The use of the ethanol pathway in goldfish Carassius auratus (L.) following anoxia

Goldfish (Carassius auratus) were subjected, for a period of 6 weeks, to 2h progressive hypoxia followed by 6h anoxia in closed respirometers at 15 degree C. The concentrations of glucose, lactate and ethanol were determined in whole goldfish following exposure to both hypoxia and anoxia. Lactate accumulation (mmol/kg/h) was 0.35 during the 1st week but declined to 0.14 in the 6th week of exposure to anoxia. In contrast, ethanol excreted to the surrounding water, increased from 65% to 92% of the total production in the lst and 6th week, respectively. The switch from lactate accumulation to ethanol pathway utilization, with the resultant metabolic depression and anoxia resistance is discussed

Efecto de la temperatura y la ración sobre el metabolismo de la carpa dorada (Carassius auratus)

[ES] Se llevan a cabo tres experimentos para estudiar el efecto de tres factores el metabolismo de Carassius auratus: temperatura de exposición, ración de alimento diario y tiempo transcurrido tras la exposición a las nuevas condiciones.

Probiotics and nutrient supplement in artificial feed of gold fish (Carassius auratus)

The efficacy of three feeds incorporated with different probiotics -Lactobacil, Vizylac and Cyfolac as nutrient supplement was evaluated in an ornamental fish, Carassius auratus (Linn.). The basal diet (40% protein) was prepared and the probiotics were incorporated at different levels viz. Lactobacil at 8x10 super(7)/100g and 12x10 super(7)/100g, Vizylac at 8x10 super(7)/100g and 12x10 super(7)/100g and Cyfolac at 12x10 super(7)/100g. Feeding trial was conducted for a period of 4 weeks. At the end of the experiment, bio-growth parameters and proximate composition of the fishes were studied. The growth, survival and protein content were improved in all the probiotic fed fishes compared to control. The maximum growth (0.74 g) and survival (85%) were observed in fishes fed with Lactobacil at 12x10 super(7)/100g. It emphasizes that supplementation of feed with probiotics has a positive impact on the growth, survival and the body composition of goldfish.

Effect of feeding bioencapsulated Lactobacillus sp. in live Tubifex sp. on the growth performance of gold fish Carassius auratus (Linnaeus, 1758)

An attempt was made to feed bioencapsulate Lactobacillus sp. in live fish food organism Tubifex for use in the culture of gold fish Carassius auratus. The C. auratus fries when fed with bioencapsulated Lactobacillus sp. in Tubifex showed significant improvement in total wet weight gain (p<0.007) and FCR (p<0.01) compared to control. The specific growth rale and mean survival were slightly higher, although insignificantly (p>0.05) in bioencapsulated Tubifex fed group. None of the bacteriological parameters of the fish gut between the experimental and control groups differed significantly (p>0.05). Lactobacillus sp. was recorded at a level of log 5.11/g on the 90th day of experimentation. When the experimental C. auratus fries were infected with Pseudomonas fluorescents, the bioencapsulated Tubifex fed group resisted the infection. The survival was significantly higher (p<0.05) in bioencapsulated Tubifex fed group (44%) than in control (22%). The C. auratus fed with bioencapsulated Tubifex showed less (55%) signs of tail/fin rot. Likewise, a significant improvement in total wet weight gain (p<0.009), FCR (p<0.01) and SGR (p<0.04) of C. auratus brooder fed with bioencapsulated Tubifex was seen compared to control group fed with depurated Tubifex.

由鲤(Cyprinus carpio)和鲫(Carassius auratus)染色体组叠加构建的两个单性人工多倍体克隆

具有天然雌核发育的多倍体杂种鱼可防止杂种优势的分离并保持其后代的杂种优势。由于假设诱发的多倍体鱼类的生殖模式是天然雌核发育的,我们进行了鲤鲫杂种的多倍体诱发,目的是描述经染色体组叠加由有性鲤鲫二倍体转化为异源三倍体及异源四倍体克隆谱系。鲤鲫杂种产生未减数而具有两亲本染色体组杂种卵子,未减数的雌核可与入卵的雄核融合叠加形成三倍体合子。鲤鲫异源三倍体胚胎发育正常,部分异源三倍体雌性个体可产生未减数的、仍保留母本的三套染色体的成熟卵子。绝大部分鲤鲫异源人工三倍体个体的成熟卵子的雌核不与入卵的雄核融合,具有天然雌

Intraperitoneal injection of extracted microcystins results in hypovolemia and hypotension in crucian carp (Carassius auratus)

Circulatory responses of crucian carp injected intraperitoneally with extracted micro-cystins (MCs) were studied at sublethal and lethal doses (150 and 600 mu g MC kg(-1) body mass, respectively). Mean arterial blood pressure (MAP), heart rate, hematocrit (Hct), red blood cell (RBC) counts, and circulating blood volume (BV) were assayed at 0, 1, 3, 12, 24, and 48 h post-toxin administration. MAP decreased significantly in a dose-dependent manner over time. Within the 48-h test period, the lethal dose as well as the sublethal dose resulted in a steady decline of MAP without recovery. Heart rate significantly increased within 24 h post-injection as blood pressure significantly dropped, then showed a terminal decline to the control level. The dose-dependent decreases in BV and Hct were directly related to the drop in MAP. Intraperitoneal injection of a lethal dose of MCs led to hepatic and gill hemorrhage. Consequently, crucian carp given MCs suffered from hypovolemic hypotensive shock. (C) 2009 Elsevier Ltd. All rights reserved.

Time-Dependent Oxidative Stress Responses of Crucian Carp (Carassius auratus) to Intraperitoneal Injection of Extracted Microcystins

This study was conducted to investigate time-dependent changes in oxidative enzymes in liver of crucian carp after intraperitoneally injection with extracted microcystins 600 and 150 mu g kg(-1) body weight. The results showed that activities of antioxidant enzymes, including superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase generally exhibited a rapid increase in early phase (1-3 h post injection), but gradually decreased afterwards (12-48 h) compared with the control, with an evident time-dependent effect. These zigzag changes over time contributed a better understanding on oxidative stress caused by microcystins in fish.

Identification and characterization of hypoxia-induced genes in Carassius auratus blastulae embryonic cells using suppression subtractive hybridization

Organisms living in water are inevitably exposed to periods of hypoxia. Environmental hypoxia has been an important stressor having manifold effects on aquatic life. Many fish species have evolved behavioral, physiological, biochemical and molecular adaptations that enable them to cope with hypoxia. However, the molecular mechanisms of hypoxia tolerance in fish, remain unknown. in this study, we used suppression subtractive hybridization to examine the differential gene expression in CAB cells (Carassius auratus blastulae embryonic cells) exposed to hypoxia for 24 h. We isolated 2100 clones and identified 211 differentially expressed genes (e-value <= 5e-3; Identity > 45%). Among the genes whose expression is modified in cells, a vast majority involved in metabolism, signal transduction, cell defense, angiogenesis, cell growth and proliferation. Twelve genes encoding for ERO1-L, p53, CPO, HO-1, MKP2, PFK-2, cystatin B, GLUT1, BTG1, TGF beta 1, PGAM1, hypothetical protein F1508, were selected and identified to be hypoxia-induced using semi-quantitive RT-PCR and real-time PCR. Among the identified genes, two open reading frames (ORFs) encoding for CaBTG1 and Cacystatin B were obtained. The deduced amino acid sequence of CaBTG1 had 94.1%, 72.8%, 72.8%, 72.8%, 68.6% identity with that of DrBTG1, HsBTG1, BtBTG1, MmBTG1 and XIBTG1. Comparison of Cacystatin B with known cystatin B, the molecules exhibited 49.5 to 76.0% identity overall. These results may provide significant information for further understanding of the adaptive mechanism by which C. auratus responds to hypoxia. (c) 2008 Elsevier Inc. All rights reserved.

Changes in plasma thyroid hormones and cortisol levels in crucian carp (Carassius auratus) exposed to the extracted microcystins

The endocrine response of crucian carp injected intraperitoneally with extracted microcystins (MC) was investigated in this study. Fish were injected intraperitoneally either with 0.75% NaCl (control) and Microcystis extract corresponding to 150 and 600 mu g microcystins per kg body weight. The plasma levels of triiodothyronine (T-3), thyroxine (T-4), free triiodothyronine (FT3), free thyroxine (FT4), and cortisol were determined at 0, 1, 3, 12, 24. and 48 h post-administration of MC-containing extract. Treated fish displayed abnormal behaviors, Such as a startle response and disoriented swimming, as well as changes in ventilation rates. Plasma cortisol concentrations of fish in both dose groups significantly increased after administration of extracted MC and remained high throughout the experiment, which suggested that MC elicited a stress response in treated fish. The profiles of cortisol changes in treated fish appeared to be dose dependent, indicating that fish in the high dose group experienced greater MC-incluced disturbance. Mortality occurred after 12 h in the high dose group. Plasma levels of T-4, T-3, FT4, and FT3 did not vary significantly between the control fish. In contrast to this, fish exposed to MC-containing extract showed significant declines in T-3, FT4, and FT3 levels in a dose-depenclent manner throughout the experiment. Plasma T4 levels, however, did not vary significantly in the low dose group, whereas they decreased significantly it 48 h post injection in the high dose group. This study demonstrates that administration of microcystins-containing extract causes a stress response and reduces the plasma levels of thyroid hormones in crucian carp. These results illustrate that microcystins exerted potent effects on the endocrine system of crucian carp, through activating their hypothalamus-pituitary- interrenal axis and disturbing thyroid function. (c) 2008 Elsevier Ltd. All rights reserved.