恩诺沙星对湘云鲫(Triploid crucian carp)组织转氨酶(GPT、GOT)活性的影响

采用胸腔注射法,研究了恩诺沙星对湘云鲫(Triploid crucian carp)组织谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性的影响。结果显示,给药后5 d,各试验组GPT、GOT酶活性均升高,与对照组之间呈显著差异水平(P<0.05);给药后10 d,各浓度组GPT酶活性有所降低,而GOT酶活性则继续升高,鳃的GOT酶活性达到了最大值;给药后15、20 d,各浓度组GPT酶活性继续下降,而肝脏、肾脏GOT酶活性在给药后15 d时达到最大值,之后开始下降;给药后25 d,鳃的GPT酶活性除200 mg/kg组外都恢复至正常水平,而肝脏、肾脏各浓度组的GPT酶活性显著低于对照组(P<0.05),肝脏的GOT酶活性恢复到了正常水平,而鳃和肾脏的GOT酶活性略高于对照组水平。

cGH Gene Transfer and PCR Detection in Crucian Carp

By inserting the CMV with HSV TK promoter into the plasmid pBlue-cGH which contained cGH gene, a chimeric expression vector was constructed for transgenic fish. After the vector was transferred into fertilized eggs of crucian carp by means of microinjection, the transgenic crucian carp was obtained. The result of PCR detection showed that the growth hormone gene of the exogenous carp was integrated into the genome of crucian carp, and the transgenic crucian carp displayed enhancement of its growth.

Effects of Salinity and Freezing on Freshness of Crucian

Salt tolerance and freeze preservation of crucian are always important concerns of people. There are rare reports on effect of different salinity on fresh- ness and quality. However, water salinity will directly affect osmotic pressure in fish, thereby affecting meat quality. In this study, the tolerance of crucian to different salinities （1‰, 2‰, 5‰, 8‰, 10‰ and 15‰） was investigated. In addition, the ef- fect of different salinity on freezing freshness and quality of crucian was studied. The results showed that the tolerance threshold of crucian to salinity was about 8‰, and the freezing quality of crucian in water with salinity of ‰ was significantly better than that in fresh water.

环丙沙星在异育银鲫(Allogynogenetic crucian)体内的积累分布及其毒性效应

采用模拟水生态系统研究了环丙沙星(Ciprofloxacin,CPFX)在异育银鲫(Allogynogenetic crucian)体内的积累和分布动态,并测定了异育银鲫体内Ⅰ相、Ⅱ相代谢酶活性的响应。结果表明,饲料暴露、水体暴露和混合暴露3种不同给药方式对CPFX在异育银鲫的体内积累趋势有明显影响,在饲料给药暴露方式中CPFX在鱼体内脏、肌肉中的残留量远高于水体给药暴露。3种不同给药方式暴露下,CPFX在异育银鲫内脏和肌肉中的积累趋势大致相同,暴露初期吸收较快,并且很快达到峰值。随暴露时间延长,CPFX在鱼体内含量逐渐下降。在CPFX暴露初期,异育银鲫体EROD酶活性受到抑制,暴露45d后,3种暴露方式异育银鲫EROD活性均恢复到暴露前水平。异育银鲫GST酶活性受到CPFX的诱导,3种不同给药方式均导致GST活性增加,并且表现出滞后响应。暴露初期不同处理组GSH含量均有下降。但随暴露时间延长,异育银鲫体内GSH受到诱导,在暴露后期又恢复至暴露前水平。

Measurement on Morphology and Kinematics of Crucian Vertebral Joints

In order to provide data for joints control of our recently designed crucian hke biomlmetlc robot fish, an A-ray photograph technology was adopted to determine the number and length of vertebral joints. A frame-by-frame analysis of high-speed videotapes was conducted to quantify the kinematics of crucian at four speeds （12.651 cm·s^-1, 18.201 cm·s^-1, 21.901 cm·s^-1, 24.368cm·s^-1） during cruising. In addition to a brief introduction to experimental conditions and methods, we analyzed the influence of individual diversity on the absolute length as well as the non-dimensional length of vertebral joints. We also presented the maximal angular velocity and acceleration of vertebral joints under four swimming speeds, and provided the change of relative rotation angle, angular difference, angular velocity and angular acceleration of the rear vertebral joints with time at a certain swimming speed of 12.651 cm·s^-1. At last, we presented the maximal lateral displacement of each mark at that speed. The study found that the influence of individual diversity on the non-dimensional length of vertebral joints is not significant; the maximal angular velocity and acceleration of vertebral joints increase with swimming speed; angular difference, angular velocity and angular acceleration exhibit two maximal values over one period at a certain swimming speed.

Comparative investigation on spindle behavior and MPF activity changes during oocyte maturation between gynogenetic and amphimictic crucian carp

The spindle behavior and MPF activity changes in the progression of oocyte maturation were investigated and compared with cytological observation and kinase assay between gynogenetic silver crucian carp and amphimictic colored crucian carp. MPF activity was measured by using histone H1 as phosphorylation substrate. There were two similar oscillatory MPF kinase activity changes during oocyte maturation in two kinds of fishes with different reproductive modes, but there existed some subtle difference between them. The subtle difference was that the first peak of MPF kinase activity was kept to a longerlasting time in the gynogenetic silver crucian carp than in the amphimictic colored crucian carp. It was suggested that the difference may be related to the spindle behavior changes, such as tripolar spindle formation and spindle rearrangement in the gynogenetic crucian carp.

Hydrodynamic Analysis of C-start in Crucian Carp

The kinematics of turning maneuvers of startled Crucian Carp (Carassius auratus) are presented. All escape responses observed are C-type fast-starts. The position of the center of mass and the moment of inertia of the fish are calculated. The results show that the position of the center of mass is always at 35% of the length of the fish from the head and the position of the center of mass and moment of inertia can be considered unchanged during C-start of Crucian Carp. Hydrodynamic analysis of the C-start is given based on the kinematics data from our experiments. The C-start consists of three stages. In stage 1, the tail fin of fish rapidly flaps in one direction, and a large moment acts on the fish′s body, which rotates around the center of mass with an angular acceleration. In stage 2, the tail fin flaps more slowly in the opposite direction at slower speed, the fish′s body rotates around the center of mass with angular deceleration and the center of mass of the fish moves along an arc. In stage 3, the moment approximately equals zero, the fish′s body stops rotating and the center of mass the moves along a straight line.

Effects of fishing handling stress on juvenile red crucian carp(Carassius auratus red variety)

The essay studies the influence of the different intensities of fishing stress on body weight （7.30 ± 1.48） g of red crucian carp juvenile body length, weight, fatness, feeding rate, specific growth rate, food conversion rate, intestinal protease, amylase and immunoglobulin. The result showed that body length, weight, fatness, feeding rate, specific growth rate of red crucian carp juvenile were significantly inhibited from fishing stress compared with the control group, and the magnitude of the inhibitory effect increased with the time going and increased. Fishing stress could increase the food conversion rate of red crucian carp juvenile, and the different intensity stress had no significant different influence. The intestinal protease activity decreased in the beginning, then increased, and was consistent with the level of the control group ultimately. High-stress group accelerated the decline of intestinal amylase activity of the juvenile. The stress increased the number of immunoglobulin of juvenile red crucian carp significantly, but the increase disappeard with the stress time extended.

miR-199-5p regulates spermiogenesis at the posttranscriptional level via targeting Tekt1 in allotriploid crucian carp

Background:Sperm abnormalities are one of the primary factors leading to male sterility,but their pathogenesis is still unclear.Although miRNAs are suggested to exert important roles in the regulation of spermatogenesis at both transcriptional and posttranscriptional levels,little is currently known regarding the regulation of sperm flagella assembly by microRNAs(miRNAs).The role of miRNAs in the development of sperm abnormalities in sterile triploid fish has not been studied.Results:In this study,we found that miR-199-5p was widely expressed in all detected tissues of different-ploidy crucian carp.As one of the testis-specific candidate markers,Tekt1 was predominantly expressed in the testis.Quantitative real-time PCR(qRT-PCR)analyses showed that the expression trend of miR-199-5p was exactly opposite to that of Tekt1.Through bioinformatics analysis,we identified a putative miR-199-5p binding site in the Tekt1 mRNA.We further identified Tekt1 as a target of miR-199-5p using luciferase reporter assay.Finally,we confirmed that miR-199-5p was necessary for sperm flagellar assembly and spermatogenesis in vivo via intraperitoneal injection of miR-199-5p antagomir or agomir in diploid red crucian carp.Moreover,miR-199-5p gainof-function could lead to spermatids apoptosis and abnormal spermatozoa structure,which is similar to that of allotriploid crucian carp.Conclusions:Our studies suggested that abnormally elevated miR-199-5p inhibited the sperm flagella formation in spermiogenesis by negatively regulating the expression of Tekt1,thereby causing sperm abnormalities of male allotriploid crucian carp.

Application Effects of Amino Acid Powder,Fermented Rapeseed Meal and Fermented Cottonseed Meal in Feeds for Allogynogenetic Crucian Carp

[Objective] To investigate the effects of amino acid powder, fermented rapeseed meal and fermented cottonseed meal on the growth performance, amino acid absorption, body composition and liver function of allogynogentic crucian carp. [ Method] The allogynogentic crucian carps with the average initial weight of 17.3 g were divided into control group and six experimental groups randomly. The allogynogentic crucian carps in the control group and experimental groups were fed a basal diet and the basal diets supplemented by amino acid powder, fermented rapeseed meal and fermented cottonseed meal, respectively. The growth rate and specific growth rate of allogynogentic crucian carps, the content of amino acids in serum and feeds, the composition indicators of body, muscle and liver, as well as the activities of glutamic-pyruvic transaminase （GPT） and glutamic-oxaloacetic transaminase （GOT） in liver and serum were determined, respectively. [ Result] The growth rate and specific growth rate （SGR） of allogynogentic crucian carps in the 3% amino acid powder group and the 6% fermented cottonseed meal group were increased greatly （ P 〈 0.05）, and no significant difference was found between experimental group and control group. The serum lysine content of the 3% amino acid powder group and the 6% fermented cottonseed meal group was higher than that of the control group. In the experimental groups, the absorption of four essential amino acids was up to a peak at 4 h after feeding, which was consistent with the control group. The body composition and liver function of allogynogentic crucian carps had no significant difference between the experimental groups and the control group. [ Conclusion] The 3% amino acid powder, 6% fermented cottonseed meal and 6% fermented rapeseed meal can substitute fish meal, cottonseed meal and rapeseed meal to improve the clrowth performance, amino acid absorption of the allogynogentic crucian carps.

Analysis of Phospholipids in Crucian Carp(Carassius auratus) Muscle by Offline HPLC-MALDI-TOF MS

The interest in the analysis of phospholipids(PLs), especially phosphatidylcholine(PC), has been increasing due to the importance of them in biochemistry as well as in industry. A method was reported based on the offline combination of MALDI-TOF MS and normal-phase HPLC for analyzing PLs extracted from crucian carp. Total PLs of crucian carp were extracted and then separated by HPLC before the collected subfractions were analyzed by MALDI-TOF MS. The mass spectra obtained show peaks of H+, Na+ and K+ adducts of PC molecules. It is shown that the prior separation of PLs by HPLC is highly necessary to remove the signal suppressing and to avoid the possible overlapping. With this method, 9 possible PC molecules in crucian carp and the corresponding fatty acid compositions were given from the well-resolved mass spectra.

Effect of lead on cytoskeletal protein stability in crucian carp Carassius auratus

Inorganic lead （Pb） is one of the most common environmental pollutants. Much evidence indicates that Pb exposure could directly affect fish growth and development. In this study, we investigated the cytotoxic effects of Pb on cytoskeletal protein stability at both protein and mRNA level in crucian carp Carassius auratus. Pb（NO3）2 treatment in concentration of 100 μmol/L resulted in decreased expression of both α- and β-tubulin but γ-tubulin as assayed with SDS-PAGE, Western Blot, and ELISA. In vivo and in vitro analyses on protein expression of tubulins are consistent. The effect of Pb on mRNA expression varied among different tissues. Our results suggest that cytotoxicity of Pb at protein translation level is stronger than at mRNA expression level.

Flash-boiled Clams with Crucian Carp

Marinate the clams in brine for 3 hours, until silt isdiffused, and keep them fresh. Draw the crucian carp,and make 3 or 4 diagonal cuts on the surface of bothsides.

Screen for stage-specific expression genes between tail bud stage and heartbeat beginning stage in embryogenesis of gynogenetic silver crucian carp

A systemic study was initiated to identify stage-specific expression genes in fish embryogenesis by using suppression subtractive hybridization (SSH) technique. In this study, we presented a preliminary result on screen for stage-specific expression genes between tail bud stage (TBS) and heartbeat beginning stage (HBS) in gynogenetic silver crucian carp (Carassius auratus gibelio). Two SSH plasmid libraries specific for TBS embryos and HBS embryos were constructed, and stage-specific expression genes were screened between the two stages. 1963 TBS positive clones and 2466 HBS positive clones were sampled to PCR amplification, and 1373 TBS and 1809 HBS PCR positive clones were selected to carry out dot blots. 169 TBS dot blot positive clones and 272 HBS dot blot positive clones were sequenced. Searching GenBank by using these nucleotide sequences indicated that most of the TBS dot blot positive clones could not be found homologous sequences in the database, while known genes were mainly detected from HBS dot blot positive clones. Of the 79 known genes, 20 were enzymes or kinases involved in important metabolism of embryonic development. Moreover, specific expressions of partial genes were further confirmed by virtual northern blots. This study is the first step for making a large attempt to study temporal and spatial control of gene expression in the gynogenetic fish embryogenesis.

Identification of serum immunoglobulin M(IgM)of crucian carp(Carassius auratus gibelio)and immune response to cyprinid herpesvirus 2 infection

Crucian carp(Carassius auratus gibelio),an extensively cultivated freshwater fish,was one of the model species for the study of fish immunology.Polyclonal antibodies were advantageous molecular tools for studying teleost immune system.Specifically,polyclonal antibodies reacting with immunoglobulins(Ig)were used successfully in studies of the teleost fishes.In the present study,we produced polyclonal antibody against CH2 domains of crucian carp IgM,and measured the in vivo dynamics of IgM mRNA caused by CyHV-2 infection.The recombinant protein IgM with relative molecular weight about 53 KD was correctly expressed in prokaryotic cells.The specificity of the polyclonal antibody was evaluated by Western blotting and results revealed that the antibody not only specifically recognized crucian carp serum but also cross-reacted with grass carp serum.Quantitative RT-PCR analysis demonstrated the expression of IgM mRNA changed significantly after CyHV-2 infection.The expression of IgM in the kidney increased and reached a maximum at 6 h post-infection(hpi),while dropped to a low level at 5 days post-infection(dpi).In conclusion,the expression of IgM was significantly upregulated in the kidney of crucian carp infected with CyHV-2,indicating that IgM played a potential role in systemic immunity against viral infection.Polyclonal antibody against crucian carp IgM had certain clinical relevance,which might provide insight into the early stage of virus infection and prevention of the disease.

Genomic variation in the hybrids of white crucian carp and red crucian carp: evidence from ribosomal DNA

In this study, we conducted a cross of white crucian carp （♀）xred crucian carp （♂） （WR）, and characterized the morphology, reproduction and genetics of the progeny. Different from parents, WR with the gray color showed the hybrid morphological traits of both parents. WR possessed normal gonads producing mature eggs or sperm, and exhibited high fertilization rate （90.2%） and high hatchery rate （80.5%）, which contributed to produce and enlarge the population. WR with the same DNA content as parents was a diploid fish with 100 chromosomes （2n=100）. Amplified ITS of 45S rDNA, in WR the sequences consisting of 884 bp bases of the entire ITS-1 region, 5.8 S region, and entire ITS-2 region. The sequences showed high similarity between WR and its parents and leaned towards male inheritance. In WR, NTS of 5S rDNA consisted of three length types with total 654 bp bases. From sequence analysis of NTS, WR shared 94.2% and 95.1% similarities with their female and male parent, respectively. Sequence analysis of ITS and NTS revealed that there existed recombination and variation in the hybrid progeny, which was the genetic base for adaptation and speciation. In conclusion, we obtained WR from hybridization and it exhibited hybrid traits in morphology and variation in genetic composition showing essential difference with its parents. The obtainment of WR has important significance in fish genetic breeding.

Biological characteristics of an improved triploid crucian carp

An improved triploid crucian carp(ITCC) was produced by crossing improved tetraploids(G1×AT,) with improved red crucian carp(IRCC,),which were obtained by distant crossing and gynogenesis.The biological characteristics of ITCC,including the number and karyotype of chromosomes,gonadad and pituitary structures,phenotype,and growth rate are reported.ITCC possessed 150 chromosomes with the karyotype 33m+51sm+33st+33t.In the breeding season,both ovary-like and testis-like gonads of ITCC were unable to produce normal mature gametes.The ultrastructure of the pituitary of ITCC showed that most of the endocrine granules in gonadotrophic hormone(GTH) cells had not been released,providing endocrinological evidence for the sterility of ITCC.Compared with triploid crucian carp(TCC) produced by mating Japanese crucian carp with allotetraploid hybrids,ITCC not only retained the excellent traits of fast growth rate and sterility,but also acquired improved morphological characteristics,including higher body,shorter tail and smaller head.

Discovery of Multiple Tetraploids in Artificially Propagated Populations of Allogynogenetic Silver Crucian Carp and Their Breeding Potentialities

1 Introduction Gynogenesis is a rare reproductive mode in fish. Among a few of species reproduced by this method, silver crucian carp (Carassius auratus gibelio) is the most special one. The specialities mainly include the following two aspects: (ⅰ)The crucian carp exists as a bisexual population which can be reproduced by natural gynogenesis, because there are both gynogenetic function and certain proportion of males in the offsprings; (ⅱ)

Discovery of Two Different Reproductive Development Modes of the Eggs of Artificial Multiple Tetraploid Allogynogenetic Silver Crucian Carp

Introduction Artificial multiple tetraploid allogynogenetic silver crucian carp possesses the reproductive function of gynogenesis in spite of incorporating one genome of common carp, because all chromosomes of silver crucian carp are maintained. This was first confirmed in the artificial propagation experiments in which the eggs of the multiple tetraploid fish were respectively inseminated with normal and UV-irradiated sperms of red common carp. Recently,

Formation and biological characterization of three new types of improved crucian carp

The improved tetraploids (G1×AT) were obtained by distant crossing and gynogenesis and the high-body individuals accounted for 2% among G1×AT. After mating with each other, the high-body in- dividuals produced three kinds of bisexual fertile diploid fishes: high-body red crucian carp, high-body fork-like-tails goldfish and gray common carp. The high-body red crucian carp mating with each other formed three types of improved crucian carp (ICC) including improved red crucian carp (IRCC), im- proved color crucian carp (ICCC) and improved gray crucian carp (IGCC). The phenotypes, chromo- some numbers, gonadal structure and fertility of the three kinds of ICC and their offspring were observed. All the three kinds of ICC possessed some improved phenotypes such as higher body, smaller head and shorter tail. The ratios of the body height to body length of IRCC, ICCC and IGCC were 0.54, 0.51 and 0.54, respectively. All of them were obviously higher than that of red crucian carp 0.41 (P<0.01). Three kinds of ICC had the same chromosome number as red crucian carp with 100 chromosomes. All the ICC possessed normal gonads producing mature eggs or sperm, which was important for the production of an improved diploid population. Compared with red crucian carp, all the ICC had stronger fertility such as higher gametes production, higher fertilization rate and higher hatchery rate. Three types of improved diploid fish population were generated from the three kinds of ICC by self-crossing, respectively. The ICC can serve as ornamental fish and edible fish. They are also ideal parents to produce triploids by mating with tetraploids. The new ICC plays an important role in biological evolution and fish genetic breeding.