• Volume 33,Issue 2,2009 Table of Contents
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    • >PAPERS
    • Comparative analysis of mtDNA gene sequences between two species of Pleuronectes

      2009, 33(2):201-207. DOI: 10.3724/SP.J.00001

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      Abstract:Pleuronectes yokohama and P. herzensteini, belonged to the family Pleuronectidae, are commercially important fisheries resources in Northwest Pacific Ocean. To date, no researches were carried out in the differentiation between the two species. Partial nucleotide sequences of mitochondrial DNA COI, Cyt b genes and D-loop were sequenced and analyzed for P. yokohama and P. herzensteini in the present day. 107 nucleotide substitutions were checked between the two species in the total 1 373 bp sequences analyzed, and most of them were synonymous transitions at the third codon positions in the two proteincoding genes. Nucleotide composition analysis indicated a strong bias against guanine (G) in the three fragments, especially in the third codon positions of COI and Cyt b gene. There was obvious difference on the degree of genetic divergence between the two species in the three mtDNA segments. The substitution rate for D-loop was the highest and the rates for COI and Cyt b were generally similar. It is suggested that gene markers used in future research should be selected carefully according to the different situations. The estimated divergence time between the two Pleuronectes species based on Cyt b gene was about 2 million years, which indicated a Pleistocene divergence for the two species.

    • The Preparation of ISCOMs with aerolysin from Aeromonas hydrophilla and the immune efficiency in Anguilla anguilla

      2009, 33(2):303-310. DOI: 10.3724/SP.J.00001

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      Abstract:In order to optimize the processes for the collection of ISCOMs, and find the immunity of aerolysin and aerolysin ISCOMs.The aerolysin gene of Aeromonas hydrophila ZN1 was orientationally inserted into the prokaryotic expression Vector pET32a(+). The recombinant plasmid was transformed into E. coli BL21(DE3). After being induced with 1mmol/L IPTG for 4h,the TrxAerolysin(Trx-AerA) fusion protein was expressed with inclusion bodies from and up to 65.5% in total recombined bacterial proteins. After being centrifuged at 40 000×g for 4 h,TrxAerolysin was divided into 5 groups for making ISCOMs: Group 1 was mixed with QuilA, Mega-10, lecithin,cholesterol,Group 2 was mixed with QuilA, Mega-10, Group 3 was mixed with QuilA,lecithin,cholesterol, Group 4 was mixed with QuilA, Group 5 only contained TrxAerolysin. All groups were centrifuged at 38 000×g for 20 h, centrifugal sedimentation was collected for analysis. After being analysed for protein concentration, SDS-PAGE and ultrastructure, the results showed that the protein yield rate of the five groups was 10.46%, 7.23%, 3.17%, 1.82%, and 0.53% respectively. The protein yield rate of the five groups had significant difference(P<0.05),Mega-10, lecithin,cholesterol improved the yield rate of ISCOMs. Next, eels were divided into 4 groups for immunization. Group 1 was injected with Trx-AerA fusion protein ISCOMs, Group 2 was injected with Trx-AerA fusion proteins, Group 3 was injected with Trx ISCOMs, and Group 4 was injected with Tris-Cl. After 30 d, the immune effects were evaluated,five eels were selected from each group for the detection of the value of sera antibody while the others were challenged with Aeromonas hydrophilla ZN1. The results showed that the RPS of the eels immunized with Trx-AerA ISCOMs,Trx ISCOMs, Trx-AerA was 83.3%(13/15),50%(9/15),and 0(3/15)after being injected with 2.0×106 CFU Aeromonas hydrophilla ZN1. The values of sera antibody to aerolysin during the group immunized with Trx-AerA ISCOMs and the group immunized with AerA were significantly higher than that of the group immunized with Trx ISCOMs(P<0.05). These results suggested that the nonspecific immune response might contribute greatly to eel immunity except in the case of sera antibody.

    • Assessing genetic diversity of the five wild tiger prawn (Penaeus monodon) populations in the coastal waters of SouthChina Sea, based on mitochondrial DNA 16S rRNA sequence

      2009, 33(2):208-214. DOI: 10.3724/SP.J.00001

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      Abstract:The aim of the present study was to survey mtDNA 16S rRNA sequence variation to assess genetic diversity and genetic structure of the wild tiger prawn in the coastal waters of South China. The samples of P. monodon had been obtained from the coastal waters of South China which included the coastal waters of Sanya(SY), Shenzhen(SZ), Zhanjiang(ZJ), Yangjiang(YJ), and Beihai(BH). 100 individuals of P. monodon were caught from the five putative populations (SY putative population, SZ putative population, YJ putative population, ZJ putative population and BH putative population). The PCR technique was used to amplify the mtDNA 16S rRNA among 100 individuals. The PCR products were purified and sequenced. As a result, 440 bp nucleotide sequences of partial 16S rRNA were obtained (some of the marginal sequences were excluded). 19 haplotypes and 28 variable sites were detected among all partial mitochondrial 16S rRNA sequences of 100 tiger prawns. Nucleotide diversities (π) in SY putative population, SZ putative population, YJ putative population, ZJ putative population and BH putative population were 0.004 35,0.005 86,0.010 50,0.010 81 and 0.011 68 respectively. Haplotype diversities (H) in SY putative population, SZ putative population, YJ putative population, ZJ putative population and BH putative population were 0.6895, 0.5211, 0.5737, 0.6000 and 0.7211 respectively. FST values between ZJ and SY,SZ putative populations, between BH and SY,SZ putative populations showed significant genetic differences, and FST values among all the other populations showed no strong genetic differences. Significant genetic differentiation among all five putative populations was found by analysis of molecular variance (AMOVA). The NJ tree of the five putative populations showed the SY and SZ putative populations had the closest relation among the five putative populations; the BH and ZJ putative populations had the greatest genetic distance from the SY and SZ putative populations among the five putative populations. Therefore, we conclude that the five putative populations can provide two basic populations for selective breeding of P. monodon; one is the SY and SZ geographic stock, and the other is the ZJ and BH geographic stock.

    • Isolation and Identification of Vibrio ichthyoenteri in Farmed Turbot and Histopathology Study of the Diseased Fish

      2009, 33(2):311-317. DOI: 10.3724/SP.J.00001

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      Abstract:In January 2007, an epizootic occurred among cultured turbot (Scophthalmus maximus) in a fish farm in Jiaonan, China. The diseased turbot exhibited hemorrhagic ulcer on the body, abundant yellowish ascetic fluid filled in the body cavity, swollen intestine, necrosis of liver and darkened cholecyst. Samples were taken aseptically from the hepatopancreas, spleen, gall bladder, kidney and brain, and inoculated immediately onto LBN (LB medium supplement with 2% NaCl) plates, which were incubated at 28 ℃ for 24 h. A predominant strain, which was designated as da3, was isolated from the gall bladder of the infected fish. Pathogenicity assays revealed that da3 was virulent to turbot by intraperitoneal injection challenge. The LD50 of da3 to turbot was calculated as 2×106 CFU/fish to the 25 g weight turbot. Strain da3 was a Gramnegative, curved bacterium with single polar flagellum, and could grow on TCBS medium and produce yellow colonies. Strain da3 was similar to Vibrio ichthyoenteri in morphological, most of the physiological, biochemical characteristics and 16S rDNA sequence except in lysinedecarboxylase (LDC) and the ability of using DGluconic acid. Strain da3 was identified to be V. ichthyoenteri. Portions of liver, spleen, kidney, intestine, brain and heart from the naturally diseased turbot were sampled, fixed in Davidson’s fixative for 12 h, and then were processed for paraffin embedding. Paraffin blocks of the tissues were cut at 4 μm thickness, and the sections were stained by the procedure of H.E. The slides were examined using an Olympus microscope, and images were captured with digital camera. There were distinctive histopathological changes in liver, kidney, spleen and intestine of the diseased turbot, and to a less extent in the brain. In the liver, necrosis of the hepatocytes were observed, and erythrocytopenia in the vein of the liver illustrated a septicemia. In the kidney, lesions of histocytes membrane, necrosis of some renal tubules, and decreased number of erythrocytes was observed. In the spleen, the number of erythrocytes decreased sharply, indicating the damage to the hematopoiesis. In the intestine, the intestinal villus goblet cells expanded to round, and the lumina popria necrosed severely. Examination of fixed tissues from healthy turbot showed structure integrity in liver, kidney, spleen and intestine. The histopathological changes in the diseased turbot may lead to the dysfunction of the organs. The results of this study may be important for controlling the bacterial disease of turbot.

    • Analysis of the similarity on ITS and 18S rDNA sequence of the main species from free-flouting Enteromorpha along China coast

      2009, 33(2):215-219. DOI: 10.3724/SP.J.00001

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      Abstract:In June and July, 2008, a large number of freefloating Enteromorpha were found in Qingdao coast, and it also happened in Lianyungang coast and Rudong coast, Jiangsu Province. The Enteromorpha samples were collected from Qingdao, Lianyungang and Rudong coasts, and their ITS and 18S rDNA genes were cloned and sequenced. Based on the sequence data of ITS and 18S rDNA, the molecular phylogenetic analysis was carried out. The result of the ITS sequence analysis showed that the similarity of ITS sequence between 5 samples (QD-01,QD-02,QD-03,LY-01,RD-02) was 100%. It indicated that main freefloating Enteromorpha species from Qingdao coast, Lianyungang coast and Rudong coast were the same species. The result was also supported by 18S rDNA sequence analysis. According to sequence similarity and BLAST analysis, there were another 2 species in freefloating Enteromorpha in Rudong. RD-01 belonged to Enteromorpha. The results of pairwise sequence divergency analysis showed that there was no difference between the main freefloating species from Qingdao, Lianyungang and Rudong coasts, and the values were very low (0.3%) between the freefloating Enteromorpha species and attached Enteromorpha species from Qingdao, Lianyungang and Rudong coasts.

    • Chemotactic response of Vibrio fluvialis to the skin mucus of Paralichthys olivaceus

      2009, 33(2):318-325. DOI: 10.3724/SP.J.00001

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      Abstract:Chemotaxis of pathogen toward the mucus layer of fish plays an important role in its colonization in the mucus. Chemotaxis is considered to be one of the virulent mechanisms of pathogenic bacterium. For a better understanding of the chemotactic response of pathogenic Vibrio fluvialis toward the skin mucus of Paralichthys olivaceus so that preventive measures can be devised to reduce the bacterial colonization, the influences of bacteria concentration, incubation time, and environmental factors, such as temperature, pH, salinity and carbohydrate, on the bacterial chemotaxis were investigated by the method of modified capillary assay and isotope tracer. The results showed that the bacteria number of chemotactic V. fluvialis toward skin mucus of P. olivaceus increased with the bacteria concentration and incubation time, respectively. Saturation of bacterial chemotaxis was obtained after incubating at room temperature for 60 min; the chemotactic bacteria increased with the incubation temperature from 5 ℃ to 15 ℃ and peaked at 15 ℃, and optimal chemotaxis was observed at pH 8; the bacterial chemotaxis weakened with the increase of NaCl concentration from 0.8 % to 3.6 %. Among 8 kinds of carbohydrate tested, mannitol, lactose and seminose promoted the bacterial chemotaxis remarkably. The results indicate that pathogenic V. fluvialis has a strong chemotaxis toward the skin mucus of P. olivaceus, and this chemotaxis is influenced remarkably by environmental factors, such as temperature, pH value, salinity and carbohydrate. The chemotaxis characteristics revealed in this paper would be helpful for the design of effective prophylactic strategies for P. olivaceus’ culture.

    • Fishing efficiency for mackerel and scad in the large light purse seine fishery estimated by generalized linear model

      2009, 33(2):220-228. DOI: 10.3724/SP.J.00001

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      Abstract:Reliable estimation of effective fishing effort, which is proportional to fishing mortality, can provide information critical to the assessment and management of fisheries resources. To estimate effective fishing effort, we need to understand fishing efficiency and factors that may influence it. In this study fishing efficiency was estimated for mackerel and scad purse seine fisheries using generalized linear model. This was done for different companies involved in the fisheries. Different choices of error structures were considered in the estimation and their impacts on the estimation were discussed. The negative binomial distribution, gamma distribution, and lognormal distribution were chosen as error distributions according to loglinear regression of variance versus logmean of CPUE (catch per unit effort). Zero CPUE values had great impacts on the assumed error structure and adding a constant (δ) to CPUE was needed for gamma distribution and lognormal distribution in maximum likelihood estimation. As δ increased, the contrast of estimated fishing efficiency was reduced greatly. Delta approaches were also chosen as an alternative way to deal with zero CPUE value in this study. Comparing the results of different models, we considered Deltanegative binomial and Deltagamma as most appropriate error distributions for this study. The results showed that the fishing efficiency differed greatly among fisheries companies and among different areas.

    • The detection of Bdellovibrio-and-like organisms in commercial preparations used for aquaculture

      2009, 33(2):326-333. DOI: 10.3724/SP.J.00001

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      Abstract:Bdellovibrio and like organisms (BALOs) are predatory bacteria and have potential for biocontrol of pathogenic bacteria. In order to provide scientific reference for study, application and market supervision of the BALOs preparations, this study was to evaluate the quality of commercial BALOs preparations used for aquaculture. Using double layer agar technique, the plaques contents of BALOs preparations manufactured by eight companies was counted with different bacteria as preys, and the prey range of plaque isolates were tested. The preparations and the plaque isolates were also identified by PCR with three pairs of taxonspecific primers targeting various BALOs groups. The 16S rRNA genes of two representative Bdellovibrionaceae isolates were sequenced and compared with that of related sequences, and the phylogenetic tree of Bdellovibrionaceae was constructed based on genetic distance analysis. The results showed that most of the BALOs preparations being examined were not qualified, only two of the eight preparations gained positive amplification by Bdellovibrionaceae primers, and showed visible plaques on the double layer agar. One of the two Bdellovibrionaceae positive preparations, JSF, had two kinds of plaque isolates with different morphologies and prey patterns, and significant differences were found when counted with different preys. About 1.4 kb 16S rRNA genes were sequenced and deposited in the GenBank with accession numbers EU884925 and EU884926 for the two kinds of plaque isolates, JSF1 and JSF2, respectively. Based on the phylogenetic analysis of 16S rRNA gene sequences, JSF1 and JSF2 were offiliated with cluster 1 (B. bacteriovorus) and cluster 5 (the species undefined) respectively in Bdellovibrionaceae. However, the plaque isolates of the other preparation contained no BALOs.

    • Research on model experiments of effects of artificial reefs on flow field

      2009, 33(2):229-236. DOI: 10.3724/SP.J.00001

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      Abstract:Effects of artificial reefs (AR) and their compounds on flow field have been studied through wind tunnel experiments under conditions of 6 m/s, 9 m/s and 12 m/s, on such single AR models with different shapes of cube, pyramid and triangular prism. As conclusions, it was found that the ranges of upwelling and wake increased with the increase of water speed; the distribution and range of upwelling and wake of the hollow models were smaller than the solid models under the same water speed, the reflux velocity of hollow models decreased with the increase of porosity of AR models; the turbulence intensity in z direction was stronger than x direction of all models. For compound cubic models, the speed of the center points increased with the increase of water speed, and the flow field has the biggest change when the distance between 1 to 1.5 times of model size at the same water speed; effects on flow field of the parallel compound models were bigger than the vertical compound models.

    • The forecasting model for incidence of ulcer disease of large yellow croaker (Pseudosciaena crocea) in marine cage culture

      2009, 33(2):334-341. DOI: 10.3724/SP.J.00001

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      Abstract:The incidence of ulcer diseases of large yellow croaker (Pseudosciaena crocea) in the cage culture was examined, and environmental factors of surrounding sea waters and weather information were surveyed in Zhoushan, during 2001-2005. Based on the data obtained, the occurrence conditions of ulcer disease and the relationship between the disease and environmental factors were investigated. The results showed that the occurrence of ulcer disease was not only related to seasons but also to the environmental factors. By means of the multivariate statistical analysis, the effects of the environmental factors on the disease were studied. The environmental factors highly related to the occurrence of ulcer disease were selected to set up models for forecasting incidence. Statistical tests showed that the forecast model was of significant efficiency for predicting the disease occurrence and the scales of diseases. The accuracy of the model reached up to 81.2% when it was used for predicting the ulcer disease of 2005 in Zhoushan. The other model was also constructed for forecasting incidence in the occurrence period in order to prevent ulcer disease timely and efficiently.

    • Relationship of crab density distribution with environment in the Oujiang estuary during summer and autunm

      2009, 33(2):237-244. DOI: 10.3724/SP.J.00001

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      Abstract:A study was conducted to define summer and autumn distribution patterns of crabs in the Oujiang Estuary and to relate these patterns to environmental variation. The result indicated that the weight densities in June and September were 112.99 and 73.48 kg/km2, respectively. Yet, the number densities were 16.84×103 ind/km2 and 3.80×103 ind/km2, respectively. In June, both of weight and number densities in the southeast studied area were higher than those in the west, and that of the south area was higher than that of the north area, in September, however, those in the east area was lower than the west. The crabs continually stayed at outer waters of investigated areas in June, especially in southeast where the water is affected by the Taiwan Warm Current, but higher density in south coastal waters in September. The low crab density in north waters of the Oujiang Estuary was ascribed to the existence of the Oujiang dilute water. Numerically Charybdis japonica was the most important crab species, but Portunus trituberculatus was a critic economical species. The hydrological factors were significantly related with weight density but not significantly with individual density. The total standing stock of crabs decreased from 50 845.5 kg in June to 33 066.0 kg in September in the whole fishing ground.

    • Degradation of Gracilaria lemaneiformis polysaccharide 

      2009, 33(2):342-347. DOI: 10.3724/SP.J.00001

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      Abstract:To discuss the effect of polysaccharide molecular weight on the antioxidation, Gracilaria lemaneiformis polysaccharide was degraded by free radical from ascorbate and hydrogen peroxide in combination, then the antioxidative activity of G. lemaneiformis polysaccharide and its degraded products was determined. The results showed that G. lemaneiformis polysaccharide was degraded basically by the reaction system of ascorbate and hydrogen peroxide in two hours, and four kinds of degraded polysaccharide were achieved with molecular size of 21 492,12 864,5 417 and 3 043 u, respectively. In the degradation, the sulfate contents were not changed but 3,6anhydrogalactose contents of degraded products decreased a little. Moreover, the higher the molecular weight of G. lemaneiformis polysaccharide, the weaker the power of reducing Fe3+, scavenging superoxide anion radical (O2-·) and 1,1-diphenyl2-picrylhydrazyl (DPPH) radical. However, the inhibitory activity againest hydroxyl radical (·OH) decreased with the degradation.

    • Histological studies on gonad of Tridentiger trigonocephalus (Gill) of the Yangtze estuary in reproductive season

      2009, 33(2):245-252. DOI: 10.3724/SP.J.00001

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      Abstract:The gonad of Tridentiger trigonocephalus (Gill) of the Yangtze Estuary in reproductive season was studied by lightmicroscope. The results showed that the reproductive season of T. trigonocephalus was April, May and June in every year. The ovaries in reproductive season were mostly in stage Ⅳ. From April to June, the ovaries in stage Ⅳ account for 62.32%, 71.43% and 48.41% of the total amount in every month respectively. The ovaries in stage Ⅲ account for 24.55%, 12.33% and 6.28% of the total amount in every month respectively. The ovaries in stage Ⅴ account for 13.13%, 10.45% and 26.67% of the total amount in every month respectively. The ovaries in stage Ⅵ account for 5.89% and 18.10% in May and June respectively. The corresponding gonadosomatic index (GSI) of ovaries from stage Ⅲ to Ⅵ were 4.49%±2.07%, 31.75%±7.08%, 38.33%±6.24% and 7.78%±3.18%. The testes were in stage Ⅳ, Ⅴand Ⅵ from April to June. The testes were mostly in stage Ⅳ in April, in stage Ⅴ in May and June. The corresponding GSI of testes in stage Ⅳ, Ⅴ and Ⅵ were 0.66%±0.07%, 0.96%±0.14% and 0.43%±0.06%. The oocytes could be divided into five phases. In the ovaries at stage Ⅲ, the oocytes were mostly at phase Ⅲ, follicular membrane and yolk grain appeared in the cytoplast. There were few oocytes at phase Ⅰ and Ⅱ at the same time. In the ovaries at stage Ⅳ, the oocytes were mostly at phase Ⅳ. The oil ball began to appear in the oocytes at the late phase Ⅳ. nucleus were excursion and distortional,zona radiate was clear at the same phase. In the ovaries at stageⅤ, the oocytes were dissociating. The two layers of follicular membrane which had outsided the plasma membrane were falling off. The ovaries at stage Ⅵ were mostly made of the oocytes at phase Ⅱ, Ⅲ and lots of discharged follicles. The testes of T. trigonocephalus were lobular type. All stages of spermatogenic cells were in the spermatogenic cysts. The wall of the spermatogenic cysts was made of sertoli cells. Spermatogenic cells in the same spermatogenic cysts developed synchronously. As spermatogenic cysts ruptured and released the sperm into lumen of seminiferous lobula after spermatozoa have matured, the developing process of spermatozoa would be completed.

    • >ROUNDUPS
    • Research progress of microsatellite DNA markers in four maricultured scallops in China: A review

      2009, 33(2):348-356. DOI: 10.3724/SP.J.00001

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      Abstract:Scallops, comprise more than 300 species identified in worldwide oceans, occupy an economically important position in many coastal countries. Until now, more than 15 scallop species have been considered as valuable fishery or aquaculture resources. Scallop adductor muscles are being considered as one of the most flavorful sea foods in markets at home and abroad, mainly due to their delicious taste and abundant nutrition. Because of the potential values, the scallop aquaculture has made great progress during the past three decades in many countries, especially in China. For examples, the farmed tideland for scallop aquaculture has exceeded 1 million hectares (ha.) and the annual production is more than 11 million tons in 2007, which makes China the firstrank not only in farmed acreage but also in annual production. However, with the blooming expansion of scallop aquaculture, some problems such as poor growth of some wild strains and largescale breakout of diseases have been arising during the past years. Further investigation and solution to these problems mainly depend on the availability of primary molecular genetic tools and the combination of efficient breeding programs with these molecular tools (i.e. molecular marker assisted selection, MAS). Microsatellite analysis based on polymerase chain reaction (PCR) offers the finest resolution for studying such genetic questions in aquacultural species. Based on the studies to date, microsatellite DNA marker has been one of the leading molecular markers for genetic analysis. Four scallops including Zhikong scallop Chlamys farreri, Noble scallop C. nobilis, Yesso scallop Patinopecten yessoensis and Bay scallop Argopecten irradians are the main species maricultured in China. In this study, we reviewed the progress in development of microsatellite DNA markers for these four scallops. We also summarized the applications of this robust molecular marker system in genetic analysis, such as genetic diversity analysis, pedigree reconstruction, genetic linkage map construction and species identification.

    • >PAPERS
    • Effects of three different incubation media on incubation length,hatching success and hatchling’s traits in yellowpond turtle(Mauremys mutica

      2009, 33(2):253-258. DOI: 10.3724/SP.J.00001

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      Abstract:Yellow pond turtle(Mauremys mutica Cantor)is a potential aquaculture target in China. Due to its poor reproduction ability, the hatching success and hatchlings quality are very important in aquaculture. In order to get some data about incubation media benefited with hatching, the effects of three different incubation media (clay, sand and vermiculite) on incubation length, hatching success and hatchling traits in yellow pond turtle have been studied in this paper. The eggs from each nest were divided into three groups and each one received one of three incubation treatments (clay, sand or vermiculite as incubation media).At room temperature, all the viable eggs were buried in media in plastic containers (315 mm×240 mm×125 mm ) that were covered with a perforated plastic membrane. During the incubation period, water would be added to the media to compensate for small evaporative losses and water absorbed by eggs. The results showed that incubation length, eggs crack ratio, hatching success and hatchling’s size (carapace length and width and plastron length and width) were affected significantly by incubation media, and some other traits of hatchlings (body height and mass and tail length) were not. The average duration of incubation in clay, sand and vermiculite was 61.2, 60.9 and 59.4 d, respectively, and the difference is significant (P< 0.05). Hatching success were 40%, 75% and 90% for clay, sand and vermiculite treatment respectively. Hatching success and eggs crack ratio were the lowest in clay, and the highest in vermiculite. Carapace width and plastron width of hatchlings from vermiculite were larger than those of hatchlings from sand and clay, and those of hatchlings from clay were the least. The locomotors performance of hatchlings from vermiculite was the best among all treatments. So, vermiculite would be the best incubation medium for yellow pond turtle. The data would be helpful to the turtle aquaculture.

    • Study on the technique of artificial reproduction and nursing of Barnea dilatata

      2009, 33(2):259-264. DOI: 10.3724/SP.J.00001

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      Abstract:The primary observation on propagation and study on artificial reproduction of Barnea dilatata were performed from 2003 to 2005.The processes of propagation and embryonic development have been found out via observation.The feasible environmental conditions for hatching,larval growth and picking of seedling were selected via experimental ecology methods. Spawning occurred naturally of Barnea dilatata in south of Zhejiang Province in April-June when water temperature is 18-22 ℃ annually.The sexually mature female individual larger than 6cm shelllength may produce more than 20 million eggs in a single spawning.The diameter of mature eggs is approximately 50 μm.The feasible temperature and salinity range 22-26 ℃ and 21-26 respectively in hatching,and the fertilized eggs develop to initial Dstage veliger for about 18 h.The feasible temperature and salinity range 22-26 ℃ and 16-26 respectively in larval growth,and the dainty food is Isochrysis galbana in D-stage larvae. After about 26-50 d rearing,the larvae develop to the stage of metamorphosis. Mud is the most adaptive substrate for metamorphous larvae to settle on.61 million juveniles of which the average shelllength was 1.43 mm were produced in the pool of 120 m2, and the density is 508 000 grain/m2.

    • Preliminary study on distribution and relative frequency of 5-HT and substance P positive cells in stomach caeca of different sizes of Onchidium strumas

      2009, 33(2):265-270. DOI: 10.3724/SP.J.00001

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      Abstract:Different sizes of Onchidium strumas were divided into three groups, which were below 8 g, 8-18 g and upward of 18 g group. The histological structure and the number of 5HT and substance P positive cells in stomachcaeca of different sizes of Onchidium strumas were observed by histology and immunohistochemical methods. The results showed that histologically, two types of villis, spindly and non-spindly, were found in stomach caeca of Onchidium strumas. The percentage of nonspindly villis was reduced with body weight of Onchidium strumas increasing. The much more spindly villis were distributed in upward of 18 g Onchidium strumas. The percentage of spindly villis (66%) was about twofold more than non-spindly villis (34%) in upward of 18 g group. Numerous 5-HT cells and substance P were detected in the epithelial mucosa of Onchidium strumas stomach caeca villis, especially in non-spindly villis. The percentage of positive non-spindly villis of both 5-HT and substance P were 10 times more than that of spindly villis. The number of positive cells of both 5-HT and substance P in stomach caeca were significantly increased in a weightdependent manner, from below 8 g to unward of 18 g group. The amount of 5-HT in upward of 18 g group (112.82 ind/cm2) was about 3fold more than that inbelow 8 g (34.8 ind/cm2), meanwhile, the amount of substance P in upward of 18 g group(177.48 ind/cm2) was about 3.5 fold more of that in below 8 g (51.08 ind/cm2). The amount of substance P was higher than that of 5-HT in three groups, and increased with weight.

    • Absorption kinetic of L-methionine and L-phenylalanine in isolated intestine of common carp

      2009, 33(2):271-277. DOI: 10.3724/SP.J.00001

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      Abstract:The experiment methods were used to determine the absorption and transportion of L-methionine and L-phenylalanine in isolated intestine.The method simulates the physiological condition in vitro. According to the change of amino acid solutions and outside the intestine that was controlled flowing through the intestine. We detemined the L-methionine and L-phenylalanine absorbed and transported across the intestine mural directly. In the experiment, the fingerling Cyprinus carpio were employed and after 24 h starvation the intestine was obtained and cultured in vitro.The concentration levels of L-methionine and L-phenylalanine were designed at 2.5 mmol/L,5.0 mmol/L,7.5 mmol/L and 10.0 mmol/L respectively. In each solution,the penicillin was added at 0.09 mg/mL,at the same time, sulfate streptomycin was added at 0.15 mg/mL.The absorbing capacity of L-methionine and L-phenylalanine in the intestine was analyse within 21 min,the concentrations of the L-methionine and L-phenylalanine in the exudation outside the intestine mural was analysed within 60 min.The result showed that the intestine of common carp could absorb and transport effectively. The mucosal influx of both Lmethionine and Lphenylalanine occurred via a carriermediated.Active transportation entry process showed MichaelisMenten kinetics.The absorption kinetic equations of L-methionine and L-phenylalanine were 1/V=0.1251×1/[S]+0.0524(R2=0.978 1,P<0.05)and 1/V=0.1887×1/[S]+0.0288(R2=0.976 2,P<0.05), respectively; At the same time, the kinetic constants of L-methionine and L-phenylalanine were Vmax=19.08 μmol/(g·min),Kmax=2.39 mmol/L;Vmax=34.72 μmol/(g·min),Kmax=6.55 mmol/L,respectively. The absorption kinetics characteristic analysis indicated that the mucosal influx and the transmural of L-methionine were higher than those of L-phenylalanine(P<0.05).

    • Molecular markers associated with body weight of mirror carp and selection and raising of progenies

      2009, 33(2):177-181. DOI: 10.3724/SP.J.00001

      Abstract (3044) HTML (0) PDF 0.00 Byte (2130) Comment (0) Favorites

      Abstract:It is the one of main directions for aquaculture breeding using molecular markers that stocks were selected by the results of quantitative traits loci of economic traits. The main goal of our long term project is to get a good variety with most economic traits in high quality by markerassisted selection. We have identified 6 microsatellite loci linked with body weight at high explained variation, and also found three of these 6 loci are homozygous in experimental population in our previous works. In this paper, using other three loci (HLJ302, HLJ338, HLJ343) which associated with body weight of mirror carp in breeding population, a group of mature mirror carp were selected and their progenies were also gotten. When the progenies grow up, the genotypes of those progenies were analyzed with these three loci. This paper compared the genotypes between 10 biggest fish and 10 smallest fish, the results showed that there are significant difference on dominant genotype linked body weight between fastgrowth population and slowgrowth population. The results showed that 10 biggest fish have the mean positive genotypes 1.7 for body weight. Meanwhile, which proved these markers are effect for breeding again. A big population with these positive dominant genotypes has been selected (also selected by good phenotypes such as good color, good body figure etc), and a big group of progenies of second generation has been produced. A new strain of fastgrowing and goodquality mirror carp has been raised.

    • The amino acids in different tissues of silver pomfret (Pampus argenteus) broodstock: Changes as the gonad development

      2009, 33(2):278-287. DOI: 10.3724/SP.J.00001

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      Abstract:The silver pomfret (Pampus argenteus) is one kind of newly exploited marine culture species in China. The nuturional characteristics of silver pomfret broodstock during the gonad development have not been reported. The amino acids of the muscle, liver and gonad tissues of the wildcaught silver pomfret (Pampus argenteus) broodstocks from Zhoushan fishery in the East China Sea were investigated with Hitachi L-8800 highspeed amino acid analyzer in this study. The changes of content and composition on amino acid in different tissues with the gonad development were also analyzed. The results indicated that the spermary in V stage of the male broodstock had the highest taurine content, 27.8 mg/g, which was significantly higher than that in muscle or in liver of the broodstocks respectively. The total amino acids of muscle, spermary and liver of the V stage of the male broodstocks were 631.4 mg/g, 544.8 mg/g and 468.0 mg/g respectively. While in female broodstocks, the taurine content (13.7 mg/g) in liver tissue was significantly higher than those in ovary and muscle tissues. The total amino acid contents of muscle and liver of the broodstocks in V stage declined significantly as the ovary developed from III stage to V stage. While the total amino acid content of ovary was stable during the ovary development from III stage to V stage. Glu, Lys, Asp, Leu and Arg were dominant in total amino acid composition of the muscle, while Glu, Lys, Asp, Val and Leu, Glu, Asp, Lys, Arg and Val, Glu, Lys, Leu, Val and Asp were dominant in total amino acid compositions of liver, spermary and ovary respectively. There were no effects of gender and ovary development stages on the total amino acid composition of the muscle. The free amino acid contents of ovaries in III, IV and V stages were 17.7 mg/g, 41.5 mg/g and 29.6 mg/g respectively. The compositions on free amino acid in ovaries were significantly variable during the development from III stage to V stage. Although the amino acids of silver pomfret broodstock at I and II stages need to be studied further, the results of this paper suggest that the total amino acid in muscle, liver and gonad and free amino acid in gonad are variable as the gonad development.

    • Estimation of genetic parameters for growth traits in rainbow trout (Oncorhynchus mykiss) with different models

      2009, 33(2):182-187. DOI: 10.3724/SP.J.00001

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      Abstract:Quantitative genetic analyses were performed to study the potential to bteed for improved body weight, body length, and condition factor in rainbow trout (Oncorhynchus mykiss) at different growth stage. In this study, samples were obtained from BaoHai Cold Water Fish Experimental Station of Heilongjiang River Fishery Research Institute, Chinese Academy of Fishery Sciences, based on 4026 hybrid individuals from the reciprocal crossing among the 5 strains between 2004 and 2006. Genetic parameters and breeding values of growth traits in rainbow trout were estimated with repeatability animal model. The fixed effects, which were included the yearseason, dam and sire strain, the permanent environment effects and the animal additive genetic effects were calculated. The phenotypic and genetic trends of growth trait in the rainbow trout were also analyzed. Growth traits were estimated utilizing Restricted Maximum Likelihood methods with a two generation pedigree. Results for heritability estimates for body length, body weight, and condition factor were 0.35, 0.10, and 0.34, respectively. The ranks of all kinds of onetrait estimated breeding values were greatly different from comprehensive estimated breeding value. And all the genetic evaluation methods also had significant correlations, which were 0.998, 0.877, 0.85, -0.071, -0.064 and -0.13. The genetic and phenotypic tendency of body length, body weight and condition factor was similar, so the selection of phenotypic value is more suitable in the study population.

    • Oncorhynchus mykiss;Oncorhynchus masou masou;hybridization; microsatellite marker; hereditary relationship

      2009, 33(2):188-195. DOI: 10.3724/SP.J.00001

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      Abstract:Interspecific hybrids of Oncorhynchus mykiss(♀) and Oncorhynchus masou masou() were produced in this study.The results showed that the fertilization rate was 90.00%, eyed egg rate was 80.52%, the hatching rate was 90.68% and the survival rate was 30.68% at fry stage.The genetic analysis on the relationship between the F1 progenies and their parents were carried out using microsatellite markers.The results are as follows: (1) Three pairs of primers only gave good images in Oncorhynchus mykiss. Ten pairs of primers gave clear bands in Oncorhynchus mykiss and Oncorhynchus masou masou, six showed specific bands and four showed no specific bands between the parents. (2) Genetic differentiation between the parents was significant at these microsatellite loci. Hybridization between Oncorhynchus mykiss and Oncorhynchus masou masou was feasible. The inherited character between parents and hybrid follows the Mendelian inheritance rule and they are indeed hybrids in genetic material. (3) The hybrid F1 has a relative genetic similarity of 0.461 7 and 0.596 5 from Oncorhynchus mykiss and Oncorhynchus masou masou, separately. The hybrid F1 has a genetic distance of 0.772 9 and 0.516 7 from Oncorhynchus mykiss and Oncorhynchus masou masou, separately. The former is farther than the later, which elucidates that the hybrid F1 has an unequal heredity difference to its parents and a genetic inclination to its paternity. This is also supported by UPGMA. The results of the research will be useful for the further study on the hybridization between Oncorhynchus mykiss and Oncorhynchus masou masou.

    • Effect of VitaminE on growth,activities of digestive enzymes of Channel Catfish,Ictalurus punctatus Rafinesque

      2009, 33(2):288-294. DOI: 10.3724/SP.J.00001

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      Abstract:The experiment was conducted to determine the effects of vitamin E on growth performance and activities of digestive enzymes of channel catfish, Ictalurus punctatus Rafinesque,and probe into mechanism tentatively. 900 healthy channel catfish [(5.00±0.50) g] of average weight were randomly divided into four groups and each group included three replicates of 75 channel catfish.Group I was supplemented with 0 IU/kg vitamin E, Other three treatments were supplemented consecutively with 50 IU/kg vitamin E for group Ⅱ,100 IU/kg vitamin E for group Ⅲ,1 000 IU/kg vitamin E for group IV.The experiment lasted for 105 days. The result showed that Vitamin E could improve survival rate,special weight growth rate(SGR),protein efficiency ratio and feed quantity of channel catfish notably (P<0.01).When vitamin E was 100 IU/kg diet, the fish had the maximal survival rate, special weight growth rate, protein efficiency ratio and feed quantity [97.78%±0.96%,2.99%±0.50%,163.71%±5.62% and (7 478.00±62.01) g/tail)], higher than the vitamin E adding amount for group 0 IU/kg extremely notably (P<0.01 ).What’s more, vitamin E had significant effect on protease, lipase in the gastrointestinal tract and hepatopancreas of channel catfish(P<0.01 or P<0.05 ). When vitamin E was 100 IU/kg diet,channel catfish had the maximal the activities of protease, lipase[(29.33%±1.66) U/(min·mg),(196.52±17.28) U/(min·mg)],higher than the vitamin E adding amount for group 0 IU/kg extremely notably (P<0.01)and higher than group 1 000 IU/kg notably(P<0.05 ). Therefore, these results indicate that vitamin E can improve the growth,activities of digestive enzymes of channel catfish at certain level.

    • Effect of dietary carnitine on growth performance, body composition and lipid metabolism enzymes of

      2009, 33(2):295-302. DOI: 10.3724/SP.J.00001

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      Abstract:An experiment was conducted to investigate the effect of dietary carnitine on growth performance, body composition and lipid metabolism enzymes of grass carp Ctenopharyngodon idella. The basal diet contained 31.36 % crude protein and 4.96 % crude fat. Five dietary treatments contained DLcarnitine concentrations of either 0(control group), 100, 200, 300 or 400 mg/kg diet. Dietary treatments were fed to apparent satiation third daily to triplicate groups of initially weighting (134.3±12.1) g per fish. Growth rate, feed conversion rate(FCR), crude fat and protein content of muscle and hepatopancreas, activities of intestine lipase, lipoprotein lipase(LPL), hepatic lipase (HL), total lipase and fatty acid synthase (FAS) were measured. At the end of the 8week feeding trial, growth rate of fish fed with 200 mg/kg carnitine diet was increased by 5.70 % and FCR was decreased by 5.26 % compared with control group(P<0.05), but no improvement was observed in other carnitine addition groups. hepatopancreas weight/ body weight, mesenteric lipid index and crude fat content of muscle and hepatopancreas were decreased(P< 0.05), but length / height and condition factor K were not affected by adding carnitine of 200-400 mg/kg diet(P>0.05). Muscle protein content was the highest at 300 mg/kg diet group. There were no significant difference in moisture and crude ash content of muscle, and protein content of hepatopancreas among all groups (P>0.05).Fish fed with 200,400 mg/kg carnitine diet, a lower concentration of serum cholesterol than the control group(P<0.05). In the respect of lipid metabolism enzymes, the addition of 200 mg/kg carnitine increased intestine lipase activity, HL activity by 28.0%, 38.6%(P<0.05), which were also increased by 41.41 %, 36.26 % by the addition of 400 mg/kg carnitine. At the same time, total lipase activity were increased by 10.66 %, and FAS activity decreased by 47.3 % by the addition of 400 mg/kg carnitine(P<0.05). Results of above show that the addition of carnitine could improve the growth performance, decrease muscle and hepatopancreas fat content, promote lipid degradation. The proper dose of carnitine in grass carp diet was suggested to be 200 mg/kg.

    • Sequence polymorphism of mtDNA D-loop in the population of the endangered species Onychostoma rara from the Qingshui River

      2009, 33(2):196-200. DOI: 10.3724/SP.J.00001

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      Abstract:The species Onychostoma rara, belonging to Cyprinidae, Cypriniformes, is endemic to China and distributed only in the Yuanjiang River and the Xijiang River systems. The species has been listed as the endangered one now. To evaluate the present state of genetic diversity of Onychostoma rara, the sequence polymorphism of mtDNA D-loop of the species is studied for the first time on the basis of determination of the 478 bp sequences from 3′ end of mtDNA D-loop in the 30 individuals of the species collected from the Qingshui River in Guizhou in Octomber, 2007, by the methods of PCR, clone and DNA sequencing. A total of 25 variable sites (about 5.23% of the total nucleotides in the sequence), including 23 transitions and 2 transitions with transversion, are detected in mtDNA D-loop of the species. The 30 individuals belong to 18 haplotypes respectively according to the determined sequences. The nucleotide diversity (π) of the 30 individuals is 0.0107, and the average number of nucleotide diffences (K) of them is 5.092. The haplotypic diversity (H) is 0.940, and the average genetic distance (P) between the haplotypes of the species is 0.014. The NJ phylogenetic tree of the 18 haplotypes comprises 2 branches. The population of Onychostoma rara in the Qingshui River includes abundant polymorphism in mtDNA D-loop sequence and high genetic diversity. The protection of population of Onychostoma rara from the Qingshui River is very necessary for the conservation and recovery of the species.

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