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。

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.

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.

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.

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 growth 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

无机的铅(Pb ) 是最普通的环境污染物质之一。许多证据显示 Pb 暴露能直接影响鱼生长和开发。在这研究，我们在鲫鱼鲤鱼 Carassius auratus 在蛋白质和 mRNA 水平在细胞骨架蛋白质稳定性上调查了 Pb 的细胞毒素的效果。Pb (在 100 μmol/L 的集中的 3)2 处理都没导致 α 导管素和 β-tubulin 的减少的表示但是是的 γ-tubulin 有 SDS 页，西方的污点，和 ELISA 的 assayed。在 vivo 并且在 vitro，导管素的蛋白质表示上的分析是一致的。mRNA 表示上的 Pb 的效果在不同纸巾之中变化了。我们的结果建议在蛋白质翻译水平的 Pb 的 cytotoxicity 比在 mRNA 表示水平强壮。

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.

Evidence for the paternal mitochondrial DNA in the crucian carp-like fish lineage with hybrid origin

In terms of taxonomic status,common carp(Cyprinus carpio,Cyprininae)and crucian carp(Carassius auratus,Cyprininae)are different species;however,in this study,a newborn homodiploid crucian carp-like fish(2n=100)(2nNCRC)lineage(F1–F3)was established from the interspecific hybridization of female common carp(2n=100)×male blunt snout bream(Megalobrama amblycephala,Cultrinae,2n=48).The phenotypes and genotypes of 2 n NCRC differed from those of its parents but were closely related to those of the existing diploid crucian carp.We further sequenced the whole mitochondrial(mt)genomes of the 2n NCRC lineage from F1 to F3.The paternal mt DNA fragments were stably embedded in the mt-genomes of F1–F3 generations of 2n NCRC to form chimeric DNA fragments.Along with this chimeric process,numerous base sites of F1–F3 generations of 2 n NCRC underwent mutations.Most of these mutation sites were consistent with the existing diploid crucian carp.Moreover,the mt DNA organization and nucleotide composition of 2n NCRC were more similar to those of the existing diploid crucian carp than those of the parents.The inheritable chimeric DNA fragments and mutant loci in the mt-genomes of different generations of 2nNCRC provided important evidence of the mt DNA change process in the newborn lineage derived from hybridization of different species.Our findings demonstrated for the first time that the paternal mt DNA were transmitted into the mt-genomes of homodiploid lineage,which provided new insights into the existence of paternal mt DNA in the mt DNA inheritance.

In vivo effects of neomycin sulfate on non-specific immunity,oxidative damage and replication of cyprinid herpesvirus 2 in crucian carp(Carassius auratus gibelio)

Neomycin belongs to the family of 2-deoxystreptamine-containing aminoglycoside antibiotics.It is widely used for bacterial infections,targeting most gram-negative and some gram-positive bacteria.Neomycin has also been reported to show antiviral activity.Here,we evaluated the toxicity of neomycin sulfate,and investigated its effect on non-specific immunity and viral infection in crucian carp(Carassius auratus gibelio).The safe concentration of neomycin sulfate for crucian carp was determined to be 102.9 mg/kg in vivo.In oxidative damage assays,neomycin sulfate increased superoxide dismutase and catalase activity and decreased malondialdehyde in the liver of crucian carp.In non-specific blood immune assays,the white blood cell count and complement 3 content significantly increased after neomycin sulfate treatment,while no significant changes were observed in antibacterial or lysozyme activity.In a challenge test,neomycin sulfate protected the crucian carp from cyprinid herpesvirus 2(CyHV-2)infection and inhibited CyHV-2 replication.In cytotoxicity assays,low concentrations of neomycin sulfate had no cytotoxicity on cells from the fins of crucian carp.The results of the present study indicate that oral administration of neomycin sulfate reduced oxidative damage,enhanced immunity and provided protection against CyHV-2 in crucian carp.

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.

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(♀)×red 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 45 S r DNA, 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 r DNA 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.

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.

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,

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)

Differential gene expression of protein kinases in oocytes between natural gynogenetic silver crucian carp and amphimictic crucian carp

A modified mRNA differential display method has been applied to studying differential expression of protein kinase genes in oocytes between natural gynogenetic silver crucian carp and amphimictic crucian carp. Total RNA was reverse transcribed using downstream 3′primers T12MA, T12MG and T12MC respectively. Then the reverse transcription products were amplified using upstream 5′kinase-specific primer designed according to protein kinase conserved sequence. The PCR products had different patterns and numbers of cDNA bands on polyacrylamide gel. Totally 21 cDNAs fragments were recovered and cloned. Two of them were confirmed to be particularly expressed in oocytes of amphimictic crucian carp, and another was specific for gynogenetic silver crucian carp.

Preliminary Confirmation of Gynogenetic Reproductive Mode in Artificial Multiple Tetraploid Ailogynogenetic Silver Crucian Carp

I Introduction Artificial multiple tetraploid allogynogenetic silver crucian carp is a special individual, which was discovered from artificially propagated populations of allogynogenetic silver crucian carp and has the obviously enhanced growth rates. One of its main specialities is that the fish maintains the whole chromosomes (162) of silver crucian carp (Carassius auratus gibelio) and fuses one chromosome set of haploid sperm (50 chromosomes ) of

Evolutionary analysis of allotetraploid hybrids of red crucian carp×common carp,based on ISSR,AFLP molecular markers and cloning of cyclins genes

The allotetraploid hybrids of red crucian carp × common carp are the first reported artificially cultured polyploid fish with bisexual fertility and stable inheritance in vertebrate. Using ISSR and AFLP markers and the cyclins genes, the genomes and cyclin gene sequence changes were analyzed between the allotetraploid hybrids and their parents. The results indicated that the allotetraploids inherited many genetic characteristics from their parents and the genetic characteristics were stable after 15 generations. However, the allotetraploids had a closer genetic relationship with their original female parents and represented a bias toward the maternal progenitor. DNA fingerprinting analysis showed that the allotetraploids had undergone sequences deletion from their original parents and that the deleted sequences were mostly from the male parent's genome. Some non-parental bands were found in the allotetraploid hybrids. Sequences analysis of the cyclin A1 and B1 genes showed nonsynonymous substitutions of single nucleotides in codons that were different from their original parents, leading to non-parental amino acid loci. We speculate that the non-additivity in the allotetraploids, compared with their progenitors, could be an adjustment to the genomic shock from heterozygosity and polyploidy, allowing maintenance of genetic stability.

Effects of 2,3,7,8-TCDD and benzo[a]pyrene on modulating vitellogenin expression in primary culture of crucian carp (Carassius auratus) hepatocytes

Vitellogenin (Vtg) is the precursor of yolk protein. Its expression and secretion are estrogen-regulated and are crucial for oocyte maturation. An in vitro xenoestro- gen screening model was established by measuring Vtg in- duction in cultured primary hepatocytes from crucian carp. Vtg production was detected by biotin-avidin sandwich ELISA method while Vtg and cytochrome P4501A1 (CYP1A1) mRNA induction were measured by semi- quan- titative PCR-primer dropping technique. Vtg and Vtg mRNA were dose-dependently induced by diethylstilbestrol (DES, 0.2—200 ng/mL) in hepatocytes of crucian carp. Co-treatment of the DES-induced hepatocytes with either 2,3,7,8-TCDD (TCDD, 0.1—4 pg/mL) or benzo[a]pyrene (B[a]P, 5—1000 ng/mL) resulted in a reduction of Vtg pro- duction and an increment of CYP1A1 mRNA expression both in a dose dependent manner, indicating the anti-estro- genic effects of the compounds. However, at lower tested concentrations, TCDD (0.1, 0.2 pg/mL), B[a]P (5 ng/mL) seemed to have a potentiating effect on Vtg expression and secretion, although by their own these compounds had no observable estrogenic effect on Vtg induction. Tamoxifen (a selective estrogen receptor modulators, 1 nmol/L—1 μmol/L), and β-naphtho-flavone (β-NF, an aryl hydrocarbon receptor inducing compounds, 2.5—1000 ng/mL) also were employed to study the possible interactions in DES-induced Vtg ex- pression. In co-treatment of the DES-induced hepatocytes with β-NF or tamoxifen, the decrease in Vtg production did parallel induction of CYP1A1 for β-NF, but tamoxifen inhib- ited Vtg induction did not parallel induced CYP1A1 expres- sion in all test concentrations. On the contrary, it was found that in co-treatment of the TCDD-induced hepatocytes with DES, TCDD induced CYP1A1 mRNA production was inhib- ited by DES also. These results implicated a possible cross talk between estrogen receptor- and aryl hydrocarbon re- ceptor-mediated pathways in the hepatocytes.