Abstract:A 155-day experiment was conducted in land-based enclosures to examine the effects of three fertilization programs on chemical water quality for integrated culture of freshwater pearl mussel Hyriopsis cumingii and fish. The pearl mussel was co-cultured with grass carp, gibel carp, silver carp and bighead carp, and the stocking ratio of fish to mussel was 1.5:1. Three fertilization programs, including fertilizing with duck manure (DM), with chemical fertilizer (CF), and with duck manure and chemical fertilizer in combination (DC), were examined. The results showed that there were no significant differences in the SD and concentration of dissolved oxygen (DO), SO42-, Cl-, CO32-, HCO3-, Ca2 , Mg2 , hardness, alkalinity, ammonia (TAN), NO3-N, NO2-N, PO4-P, total nitrogen (TN), total phosphorus (TP) and chemical oxygen demand (CODMn) between the fertilization treatments. The ratio of PO4-P to TP was higher in the CF enclosures than in the DM enclosures. The pH was lower in the CF enclosures than in the DM enclosures during the earlier phase (from May 20 to July 18). With the progress of the experiment, the SD, concentration of DO and Ca2 , and ratio of Ca2 to Mg2 decreased, while the concentration of TAN, NO2-N, TN, TP and CODMn increased. The results of this study indicate that using different fertilization programs had no significantly different effects on the major ions, hardness, alkalinity, TAN, NO3-N, NO2-N, PO4-P, TN, TP and CODMn in the enclosures with integrated culture of freshwater pearl mussel with fish and fed formulated fish feed intensively. However, the concentration of CODMn was lower, but the ratio of PO4-P to TP was higher, in the enclosures fertilized with chemical fertilizer than in the enclosures fertilized with duck manure. The concentration of CODMn and ratio of PO4-P to TP were at median levels in the enclosures fertilized with chemical fertilizers and duck manure in combination. The higher growth rate and nacre secretion of H. cumingii hung in the enclosures supplied with both chemical fertilizers and duck manure suggests the water chemistry parameters in these enclosures are suitable for H. cumingii farming.

Abstract:In order to analyze the seasonal variation characteristics of emissions of CO2 ,CH4 , N2O and evaluate the comprehensive global warming potentials (GWPs),Fluxes of CO2,CH4 and N2O were measured simultaneously from the culture ponds of Chinese mitten crabs by the closed static opaque chamber-GC techniques. The results showed that the pond ecosystem were the sources of CO2

Abstract:The mature spawners selected from Japanese flounder selection and breeding founder were used to produce the normal diploids (ND), meiotic gynogenetic diploids (MGD-1), successive two generation meiotic gynogenetic diploids (MGD-2) and inbred diploid (MGD1H) by crossing mature female from MGD-1 with induced sex reversed male. A set of 72 microsatellite markers distributed evenly on 24 linkage groups were selected from the published genetic linkage maps of Japanese flounder. To estimate microsatellite-centromere map distances, four MGD-1 lines were produced. Under the assumption of complete interference, 17 markers were located in the centromeric region, 36 in the distant region from the centromere and 19 in the intermediate region of linkage groups. The genetic characteristics of four types of diploid in Japanese flounder were identified by these markers. Analysis showed that the number of allele (A) and polymorphism information content (PIC) have limited variation range, in which, the A and PIC of ND were the highest while those of MGD1H were the lowest. In four types of diploid, the observed heterozygosity decreased and homozygosity increased as microsatellite-centromere map distance increased. The highest and lowest percentages of homozygous individuals were found in centromeric region from 8.8% to 29.1% and in telomeric region from 2.4% to 23.2%, respectively. In which, the variation range of MGD-1and MGD-2 were significant higher than that of the rest diploids. Thus, choice of microsatellite markers located on different positions in linkage group have greatly impact on identifying genetic characteristic of different diploids in Japanese flounder.

Abstract:In order to elucidate the dynamics of fishery resources structure in the Yellow Sea, based on the bottom trawl survey data by R/V “Beidou” from September, 2006 to May, 2007 in the sections of the Yellow Sea (S1 section locates in the northern part of the southern Yellow Sea, S2 section lies on the border of Yellow Sea and East China Sea, S3 section locates in the central part of the southern Yellow Sea), the species shifts, abundance dynamics, fishery species diversity and their relative factors were analyzed. A total of 156 species were collected, mainly composed of fish species (109 species), including 97 demersal species; the top ten species catch were beyond 60% of the total catch, and the species composition were similar, as well as the species composition in every section (excluding S2 section), including Larimichthys polyactis, Lophius litulon, Liparis tanakae, Engraulis japonicus, Crangon affinis and so on, but the proportion of these species in the total catch were greatly changed. The biomass spectra in every section mainly distributed not beyond 40g. The diversity of fishery species decreased in each section from the northern part to the southern part of the southern Yellow Sea, and the diversity of fishery species was positively correlated with species richness. From above mentioned, the significant seasonal changes were found in the fishery resource structure of the sections of the southern Yellow Sea, these changes mainly attributed to the variations in the relative abundance of the main fishery species, also closely related to the sea bottom temperature and sea salinity.

Abstract:Midkine is a kind of heparin-binding cytokine, and it promotes growth, survival, migration and other activities of target cells. The expression of fusion protein MK will facilitate further studies for its biological functions in half-smooth tongue sole (Cynoglossus semilaevis). Based on the plasmid containing full length of half-smooth tongue sole MK cDNA sequence, the site-directed mutagenesis of predicted mature MK sequence was successfully obtained by SOEing PCR method according to the E. coli codon bias. After being confirmed by sequencing, the resulting gene was cloned to the vector pET32a ( ) to yield an identified recombinant plasmid pET32a( )-MK, which was then transformed into E. coli host strain BL21(DE3)pLysS. SDS-PAGE analysis revealed that high level of fusion protein MK was induced by 1mmol/L IPTG at 37℃ and was mainly detected in sonicated supernatant. The yielded fusion protein MK was approximately 33 kD which was in accordance with the theoretical molecular weight. This research establish the bases for further study for its detailed functions in half-smooth tongue sole.

Abstract:In each spring during 1989-1993 and 2003-2012, oyster samples（Crassotrea rivularis Gould）were collected from 16 sites along Guangdong coast, China. The contents of DDTs （including o,p’-DDT，p,p’-DDT，p,p’-DDD，p,p’-DDE) were detected by Gas chromatography with electron capture detector. Based upon the detected data of DDTs in oyster along Guangdong coast and the data taken from the Pollution Investigations of Guangdong coastal zone, China during 1980-1985, the residues levels, spatial and temporal trends and compositions of DDTs in oysters were analyzed and discussed. Also, food safety and biological quality of DDTs in oysters were assessed. The results indicated that DDTs was detected in 95.9% of the samples, and in the detectable samples, DDTs contents ranged from 0.11 to 76.3 ng/g with an average of 3.87 ng/g on wet weight basis. DDTs contents decreased obviously during the early period after DDTs was banned from 1980s to early 1990s, but presented a gentle increase owing to possible inputs of new DDTs from 2003 to 2007, with the national implementation measures for the Stockholm Convention, DDTs contents maintained steady finally from 2008 to 2012. In early 1980s, DDTs contents in Pearl River Estuary were much higher than East Guangdong coast and West Guangdong coast, but the difference of DDTs contents from these three coasts narrowed gradually with time and became not significant statistically since 1989（P >0.05）. The composition analysis of DDTs in the oyster samples showed that DDTs residues were sourced mainly from the historical usage, but the higher ratios of DDT/ (DDD DDE) and o,p’-DDT /p,p’-DDT（>1）indicated that there was probably input of new DDTs which might come from dicofol in periods of 1989-1993 and 2003-2007. The DDTs residues levels of all oyster samples were below the state residual limited standards, thought during the period of 1980s-1990s and 2003-2007, there was 4.6% of oyster samples exceeded US EPA reference limits(14.4 ng/g), and 6.7% of samples exceeded the requirement of the first grade of the National Marine Biological Quality Standard of China（10 ng/g）, the biological quality of DDTs in oysters was good and the food safety was acceptable in recent years.

Abstract:Swordtail fish, Xiphophorus helleri, is an ideal experimental animal for Ecological toxicology studies, which is sensitive to environmental contaminants. The construction of related cell lines can provide foundation for its application. The aim of this study were to establish the swordtail fish brain cell line SFB (swordtail fish brain cell line) and to study its CYP1A mRNA inductive effect. Primary brain cell culture of Swordtail fish was initiated by trypsin digestion methods, and a stable brain cell line was obtained successfully after culture and proliferation. The optimal culture medium for SFB was Leibovitz-15 to blend DMEM/F12 medium with 1:1 supplemented with 15% fetal bovine serum, and the optimal culture conditions for SFB were 27℃ and 5% CO2. The SFB grew actively under the optimal medium and culture conditions, the doubling time is 43. 048 hours at 65th passage . And chromosome analysis showed that the SFB exhibited chromosomal a diploidy with a modal chromosome number of 48, the karyotype formula was 2n=2st 46t, NF=48, which consistent with the swordtail fish.The cells maintained their original shape and high viability after cryopreserved and resuscitated. The induction experimental results showed that, the expression of CYP1A mRNA in B(a)P-treated (10-8~10-5mol/L) SFB cells increased significantly, and increased along with the increasing dosage of B(a)P. A dose-dependent relationship between the levels of CYP1A mRNA expression and doses of B(a)P was observed. Thus, the establishment of this cell line will provide convenience for the toxicological evaluation and a basis for its application in Ecological toxicology.

Abstract:Accumulation and elimination of semicarbazide (SEM) in adductor, mantle and gill, digestive diverticula of Chlamys farreri under laboratory conditions were studied using ultra performance liquid chromatogram tandem mass spectrometry in this work. The Chlamys farreri was divided randomly, then exposed to seawater containing 1.0 μg/L, 5.0 μg/L and 20.0 μg/L SEM for 8 d at (14±2)℃. After exposure, Chlamys farreri was placed into clean seawater for the subsequent elimination experiment. As the increase in the concentration of SEM in seawater, the accumulation amount in three various tissues was increased. The digestive diverticula has the maximum of the biggest concentration (Cmax), average elimination speed and the greatest accumulation ability of SEM. The accumulation capability of the various tissues was as follows: digestive diverticula>mantle and gill>adductor; and the accumulation amount in the three tissues increased with the addition of SEM concentration in seawater, presenting significantly positive correlation. SEM was accumulated mainly in digestive diverticula and mantle and gill, and the content of SEM in adductor was relatively lower. The elimination capability of the various tissues was as follows: adductor>mantle and gill>digestive diverticula; but the SEM content in adductor could be reduced to less than 0.50 μg/kg after a period of purification at higher concentration. DAS 2.0 pharmacokinetic parameter calculation program was used to process SEM content-time datas and to calculate the relevant pharmacokinetic parameters. The results indicated that area under concentration-time curve (AUC) differed greatly in the three tissues and the tissues had different accumulation capability. For example, in the concentration of 20.0 μg/L, the content of SEM in adductor,mantle and gill,digestive diverticula was 5.94 μg/kg, 9.87 μg/kg and 15.81 μg/kg respectively after 42 d elimination experiment, which were also much higher than the specified values. When Chlamys farreri was bred in clean seawater, the SEM accumulated in the body was eliminated significantly. The excretion rate in adductor,mantle and gill,digestive diverticula was 91.3%、90.0%、89.6% and gradually reduced. Under the experimental conditions, when Chlamys farreri exposed to seawater containing 1.0 μg/L, 5.0 μg/L and 20.0 μg/L SEM for 8 d at (14±2)℃, it was recommended withdrawal period of 15 d, 50 d and 70 d respectively. When the water temperature was highter than the experimental conditions, the withdrawal period could be shorten; and extended in lower water temperature. The accumulation and elimination curves of SEM in Chlamys farreri provided useful information for studying decontamination of seashells that have SEM contamination.

Abstract:The mitochondrial cytochrome b gene fragments of eight cyprinoid aquaculture breeds were sequenced and their genetic diversity and cluster analysis were compared. Mean content of T, C, A, G, are 26.8﹪, 29﹪, 29.7﹪, 14.5﹪, respectively, and A plus T is higher than G plus C. The mutations in the 141 amino acids translated by 425bp of Cyt b gene fragment, could not affect its function dueing that the majority were generated between the valin and isoleucine which were unchanged and hydrophobic. The result demonstrated that the evolution of this protein was slowly, and it was adapted for the genetic analysis. Five haplotypes were detected from 140 individuals, of which 75.5 percents belong to the Hap2, and the Hap1 was mainly distributed in the mirror carp. The AMOVA and cluster analysis based on Kimura 2-parameter model exhibited very close relationship among Germany mirror carp, Scattered mirror carp, Red mirror carp, and also revealed that the relationship among Cyprinus carpio blue var, Cold-resistance strain of red purse carp, Frigid carp was close. The results were in accordance with their breeding backgrounds.

Abstract:Grass carp reovirus (GCRV) has been considered as the most pathogenic agent and poses a significant threat to grass carp culture. A virulent reovirus strain, HZ08, was isolated from diseased grass carp in Zhejiang province, and has been proven to be epidemic strain in China. To development serological methods for detecting prevalent GCRV, the gene encoded for major outer capsid VP4 was selected clone to plasmid pET32a( ) and the purified recombinant VP4 protein was injected into BALB/c mice through subcutaneous route. The spleen cells from immuned BALB/c mice were fused with SP2/0 myeloma cells, and three hybridoma cell lines, designated as 2C2, 2F3 and 5E5 respectively, were screened out to be able to secret monoclonal antibody (McAb) against VP4 protein using indirect ELISA. All the McAb were IgG1 subtype with κlight chain and could not react with GSRV, ISKNV, IHNV except GCRV-HZ08. The hybridoma cell line 2C2 was selected for McAb preparation, and the titers in cell culture medium of the ascetic fluids were up to 1:720,000. The results of western blot and IFA showed that the McAb could recognize the authenticVP4 protein of GCRV HZ08 particles. In present study, the McAb against VP4 of GCRV HZ08 was successfully prepared, which laid a foundation of developing a rapid determination method for GCRV and further study of structures and functions of VP4 protein.

Abstract:Morphological and genetic differences of three Sinibrama macrops populations from Qiandaohu Lake were compared by traditional morphological method and mitochondrial D-loop region sequence analysis. Based on the results of principal components analysis, specimens clustered together by scatter plots of scores could not be divided. It indicated that there is no morphological differentiation among all samples from Qiandaohu Lake. 27 polymorphic sites were detected yielding 72 haplotypes. NJ tree and network showed that the geographic distribution was ambiguous. There were similar base composition among the three populations, and the content of A T (64.82%) was higher than the content of G C (35.18%). High genetic variation (h) and low nucleotide diversity (π) were found in the three populations analyzed for D-loop, and the diversity of Fenkou (h=0.968?0.018, π=0.00476?0.00049) was the highest, Linqi (h=0.968?0.017, π=0.00453?0.00041) was the second, Fuwen (h=0.937?0.037, π=0.00330?0.00035) was the lowest. The values of genetic differentiation index (Fst) were lower than 0.05 and not significant (P>0.05). A strong gene flow among the populations were observed (Nm>4), and the Nm between Fenkou and Fuwen was highest (Nm= 27.40). The results of all supported the three geographical populations belong to a randomly mating population. Consequently, we suggest to keep the pattern of gene flow of S. macrops, and reduce the influence of human activities so as to protect this important species in fishery resources.

Abstract:This experiment was conducted to study the effects of complex probiotics on water quality and microbial community structure in grass carp culture. The results of water quality determination showed that compared with the control group, the concentration of ammonia nitrogen, nitrite nitrogen and total nitrogen in treatment group was lower in the whole experiment, respectively; At 18th d, the concentration of nitrate nitrogen in treatment group was decreased by 54.49% (P<0.05). At 15th d, the concentration of total inorganic nitrogen in treatment group was decreased by 28.75%（P<0.05）. Orthophosphate and total phosphorus was no difference between the two groups. The 454-Pyrosequencing results showed that the composition of bacterial communities was highly variable between control and treatment group, and microbial diversity of treatment was higher than that of contorl. Firmicutes and Proteobacteria was decreased by 91.21% (P<0.01) and 21.75% (P<0.01) respectively. Bacteroidetes, Actinobacteria and Cyanobacteria was increased by 2.88 times (P<0.01), 4.35 times (P<0.01) and 8.48 times (P<0.01) respectively. Among Proteobacteria，Alphaproteobacteria and Betaproteobacteria was elevated by 3.18 times (P<0.01) and 18% respectively, when Gammaproteobacteria was reduced by 78.82% (P<0.01).

Abstract:Basing on one generation of individual selective breeding and four generations of family selective breeding of Litopenaeus vannamei, 120 selective breeding families of resistance to WSSV were produced from 2002 to 2007. Experimental WSSV challenging tests showed that the survival rates of generation 2(G2)，G3，G4 and G5 were 5.57±9.83%, 8.66±11.52%, 9.52±8.84% and 13.79±12.86%0%, and the variation coefficient of survival rates were 1.77,1.40,0.97and 0.87, respectively. Each generation of selective breeding families of resistance to WSSV were divided into three groups: relatively high resistant families, mid-resistant families and susceptible families post WSSV infection. The proportion of susceptible families in G2 to G5 were 76.5%, 55.2%, 51.4% and 33.3% ,and the survival rate were 0.44 ± 1.09%,0.78 ± 1.70%,2.27 ± 2.76% and 2.44± 3.09 %,respectively ,with a acute mortality peak appeared at 2nd ~3rd day post infection. The proportion of mid-resistant families in G2 to G5 were 0、20.7%、31.1% and 38.5% ,and the survival rate were 0、9.08±1.46%、10.7±1.41%and 11.36±3.30% ,respectively ,with two mortality peaks appeared at post WSSV infection, and the 1st peak value was greater than that of the 2nd peak. The proportion of relatively high resistant families in G2 to G5 were 23.5%、24.1%、17.1% and 28.2% ,and the survival rate were 22.23±5.21%、22.7±12.30%、24.45±6.56% and 28.98±8.09% ,respectively ,with two mortality peaks appeared at post WSSV infection, and the 1st peak value was less than that of the 2nd peak, which showed that these families in this group possessed strong resistance to WSSV infection. So the eruptible time of WSSV was delayed and the valuable opportunity could be offered to take effective measure to prevent and cure. However, the average of spawning quantity of each generation was declining (p <0.05) year after year.

Abstract:Oscillatoria sp. is a so common and harmful cyanophytes in shrimp culture pond. In this experiment, sterilization filtrate of Brevibacillus laterosporus and Oscillatoria sp.were used to study algicidal effects in their different growth stages. Algae dry weight, chlorophyll-a and phycocyanin protein contents were measured in order to explore the best algae-lysing period and algicidal mechanism. The final aim is that providing theoretical basic for improving water envinment in shrimp pond. The results show that algicidal effects of sterilization filtrate to Oscillatoria sp. are very significant(P<0.01)when the algae-lysing bacteria in the stable and decline phases. After 7 days, algae dry weight were reduced by 51.77%、47.04% separately, chlorophyll-a content were decreased by 67.60% , 59.13% respectively, phycocyanin protein content were severally increased by 33.51%、30.97%. And similarly, the algicidal effect of sterilization filtrate of alage-lysing bacteria is so conspicuous(P<0.01)to the lag phase Oscillatoria sp.. 7 days later, algae dry weight were reduced by 63.90%, chlorophyll-a content were decreased by 69.72%, phycocyanin protein content were increased by 54.17%. The conclusion is that sterilization filtrate of Brevibacillus laterosporus in the stable phase has the best algicidal effect to the lag phase Oscillatoria sp..

Abstract:Sex hormones are involved in animal sex, gonad development and reproductive cycle regulation and other functions, it"s known that 17beta-hydroxysteroid dehydrogenase (17β-HSD4) had played an important role in synthesis and metabolism of the sex hormone. 17β-HSD4 mainly catalyzes to deactivate the biological activity of estradiol and androstenediol. In the present study, we got a partial expression sequence tag of 17β-HSD4 from the transcription group database of Chlamys farreri firstly, then a cDNA was cloned by using Rapid Amplification of cDNA End (RACE) methods, of which the full-length is 2217 bp, contains an open reading frame of 2223 bp and encodes a peptide of 740 amino acids. Homology analysis and phylogenetic tree revealed that it"s 17β-HSD4. The mRNA was detected widely in the testis, ovary, adductor muscle, mantle, gill, hepatopancreas and kidney, especially higher in hepatopancreas and kidney. The result of qRT-PCR showed the expression of 17β-HSD4 gene in testis at mature stage is obviously higher than the testis at proliferative stage and resting stage, and ovary of all stages. It could be concluded that 17β-HSD4 can be synthetized in many tissues of Chlamys farreri, also might regulate the maturity of testis.

Abstract:Microbial flocs is a complex of bacteria, residue of feed, algae, protozoa in aquaculture. Biofloc technology (BFT) based on microbial flocs have been applied in the culture of Oreochromis mossambicus, Litopenaeus vannamei, Macrobrachium rosenbergii and so on. In this study, microbial flocs was achieved by artificial culture in natural marine water. In order to investigate the active ingredients in the microbial flocs, molecular weight, content of total polysaccharide and amino acid in the microbial flocs were analyzed by methods of gel permeation chromatography, Phenol sulfuric acid colorimetry and automatic analyzer for amino acid respectively. The results of preliminary biochemical analysis showed that the average molecular weight of extractive from supernatant of microbial flocs was about 213281Da, the polysaccharide and amino acid content of sediment from microbial flocs was 29.57% and 12.63% respectively. Further study was conducted to investigate the function of microbial flocs to enhance the immune activity of shrimp. Microbial flocs were added in the Litopenaeus vannamei diet at a rate of 0%, 0.02%, 0.1%, 0.5%, and 2.5%. Activity of antibacterial, lysozyme and phenoloxidase (PO) in serum of shrimp were analyzed after 14 days of feeding trial. Challenge test was carried out to evaluate the effect of microbial flocs on improving the ability of shrimp against virulent bacterium of Vibrio Harveyi after 30 days of feeding trial. Results show that activity of antibacterial and lysozyme in serum of shrimp fed diet containing microbial flocs at a rate 2.5% were higher than that in other groups (p<0.05). PO activity in serum of shrimp fed diet containing microbial flocs at rates 0.5% and 2.5% were higher than that in other groups(p<0.05). Challenge test show that cumulative mortality of shrimp fed diet containing microbial flocs at a rate of 0.1% was the lowest than that in other groups(p<0.05). The present results suggested that microbial flocs can be utilized by shrimp and can improve the ability of non-specific immunity of shrimp.

Abstract:The Y-box protein family is a highly conserved nucleic acid binding proteins which are conserved from bacteria to human. The new member of this family from Strongylocentyotus intermedius were isolated and characterized. The full-length sea urchin Y-box cDNA was 2425bp in length and contained a 996bp open reading frame (ORF), a 1348 bp 3 UTR and a 81 bp 5 UTR. The full-length Y-box cDNA had AATAA special sequences in the 3′ UTR and polyA regions. The full-length Y-box cDNA encoded 331 amino acids constituting a 37.4KD protein molecule with an isoelectric point of 8.55. The precursor protein was hydrophily. The Y-box protein contain three protein domain involved N-glycosylation site, hydrophily C-tail end and cold shock domain CSD, without transmembrane. CSD area in Y-box protein family was very conservative .The secondary structure of the precursor protein consisted primarily of a 1.98% alpha helix, with a 9.97% strand chain and random coil 88.22%, without β folding. The comparison revealed that the similarity of the Y-box precursor protein ranged from 22.37% to 41.26% with other animals. Y-box protein secondary structure is very similar with human and zebrafish. The evolutionary tree indicated that the Y-box precursor protein was clustered with Aplysia californica、Chlamys farreri and Ciona intestinalis. Xenopus tropicalis、Gallus gallus、Mus musculus、Homo sapiens、Lethenteron camtschaticum and Carassius auratus belong to vertebrates. Chironomus tentans and Drosophila virilis belong to land vertebrates. The Y-box gene expression pattern was detected in coelomic fluid, tube feet, peristomial membrane, male gonads, female gonad, muscle and intestinal tissue. The results in sea urchin Strongylocentrotus intermedius indicated that the difference was not significant. The expression of Y-box gene under temperature stress was reduced, and the expression quantity of Y-box gene decreased gradually with the increase of temperature. The results provide a reference for investigating the function of this gene.

Abstract:Gynogenetic Orange ornamental carp(Cyprinus carpio L.,Koi) with high hatch rate and survival rate was obtained by egg activation using the UV-inactivated sperm form Blunt snout bream(Megalobrama amblycephala) and then chromosomes doubling with cold shock treatment at the temperture of 4℃ for 30min. The gynogenetic Orange ornamental was proved to be diploid with 100 chromosomes by chromosome number detection and karyotype analysis. At the age of 1 year old, 10 gynogenetic fish were randomly sampled to detect gonad development and the only ovary was observed in all the samples; during the spawning season at the age of 2 years old, 100 gynogenetic fish were striped for semen production but none of the sample could strip out semen but green eggs, the presence of all-female gyongenetic Orange ornamental carp provided vital evidences to prove that the sex determination type of female Orange ornamental carp is XX. In addition, the genetic relationship of SOX gene, the phenotypes and body color between gyongenetic Orange ornamental carp and its parent were also observed in this study, the results show that gyongenetic Orange ornamental carp inherits the same amplified fragments as the maternal parent, which provide importment evidences to get rid of its heterozygosity. Meanwhile, gynogenetic Orange ornamental carp inherited the graceful shape of its maternal parent but the body color is brighter. The formation of gynogenetic Orange ornamental carp lay foundation on purification，rejuvenation and genetic improvement for Orange ornamental carp and also provided good material for fish sex determination and sex control research.

Abstract:The paper studied the growth and development of sexual characters, primary sexual maturity, sexual maturity of pond-reared Penaeus monodon in relation to culture conditions in several years. The results showed that the carapace length was positively and linearly correlated to the pudendum length and thelycum width of pond-reared P. monodon. The minimal average carapace length, body length and body weight at primary sexual maturity of pond-reared males and females were 3.1 cm, 11.1 cm,20.0 g and 4.3 cm, 15.1 cm, 53.0 g, respectively. There were not different among different culture conditions. The average carapace length, body length and body weight at sexual maturity of pond-reared males and females were 3.7 cm, 13.0 cm, 37.0 g and 5.0 cm, 17.0 cm, 75.0 g, respectively. The sexual maturity was positively and linearly correlated to day age of pond-reared Penaeus monodon. The culture conditions (density, natural food) were key factors known to influence the growth, development and sexual maturity of pond-reared Penaeus monodon. The earliest time of spermatophore production of the tide pond males (120 days) was significantly earlier (P < 0.001) than the earthen pond males (150 days). The day ages of sexual maturity of male were 160 days (tidal pond) and 260 days (earthen pond), respectively. Prawns started mating at 165 days from tidal pond and at 240-280 days from earthen pond, reached full ovarian maturation and spawned as early as about 205-236 days (tidal pond) and 295-360 days (earthen pond).