• Volume 33,Issue 4,2009 Table of Contents
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    • Morphological development and growth characteristics of the juvenile of Hyriopsis cumingii

      2009, 33(4):604-609. DOI: 10.3724/SP.J.00001

      Abstract (3827) HTML (0) PDF 0.00 Byte (1805) Comment (0) Favorites

      Abstract:The morphological development of the juvenile of Hyriopsis cumingii was observed and described from the glochidia that fell off the host fish to the triangle in a hatchery at the water temperature of 26.5-32.0 ℃. According to the morphological characteristics, the juvenile development of Hyriopsis cumingii could be divided into four stages. The average shell length of newly fallen juvenile was 221.88 μm, and the sail had not been formed, so its total height was equal to shell height. The width of the shell was expanding quickly in the first four days after falling off, named as width expanding stage. From fifth to twentythird days, the juvenile developed into umbo extrudeing stage, because the umbo began to extrude. There are apparent changes during this stage that the posterior shell was much longer than the anterior shell after fifteen days. On twentythird day, the wing appeared, so the juvenile began to develope into wing forming stage. From thirtyfourth to sixtieth day, the sail gradually grew and the ratio of total height to shell height increased. The umbo was no longer the top of the juvenile and it developed into sail growth stage. On sixtieth day, the development of the juvenile stage of Hyriopsis cumingii had completed and entered the young stage, with similar morphological characters to those of adult. The position of the umbo represented the most apparent changes during the juvenile development of Hyriopsis cumingii. The relationship between total length and day age could be expressed as 〖L=329.39e0.059tr=0.968), that between total height and shell length as 〖H=0.776L-103.36(r=0.997).

    • >ROUNDUPS
    • Progress on sexual selection researches of aquatic animals

      2009, 33(4):704-712. DOI: 10.3724/SP.J.00001

      Abstract (3891) HTML (0) PDF 0.00 Byte (1867) Comment (0) Favorites

      Abstract:Sexual selection was referred to as the mate choice between males and females, and played an important role in the process of speciation and species evolution. The past two decades have seen extensive growth of sexual seletion researches. Though most of these researches were focused on terraneous animals, studies on aquatic animals were gradually increasing, especially for fishes, sea urchins, and oysters, etc. Unlike most of terraneous animals, the sexual selection of aquatic animals is original and conservative, for example, no perfect sense organs, no remarkable genitalia and nearly no parent behavior. However, through a long time evolution process, the mode of sexual selection in some aquatic animals is also unique. To summarize recent research results and enlighten novel research ideas of sexual selection, here, we have a review on hydrobios sexual selection. The sexual selection modes are various in aquatic animals, and change from individual to cell levels. Like high terraneous animals, there are conspicuous sexual dimorphism between male and female in some fishes, such as Xiphophorus helleri and Goodea atripinnis. Additionally, body color, body size and even aggressive reaction of males are all seletive objects for females in the mate choice. However, unlike terraneous animals, quite a number of sexual selection exist between sperms and eggs, such as sea urchins and oysters. The genes coding bindins and bindin receptors are high polymorphism in sea urchins, and it was believed that the polymorphism was the selection basis of eggs for sperms. Additionally, there is the phenomenon of sex role reverse in some fishes, such as pipefish and sea horse, which is similar to some insects. What is the mechanism of sexual selection in aquatic animals? At present, many researches have given stronger supports that the mating selections were mainly based on both of additive genetic benefits and nonadditive genetic benefits. It is worth noting though most of sexual selection is benefitial to the fitness of aquatic anmials, some mating choices are deleterious, for example, the males attracted to females yielded more low viability offspring than other males in Poecilia reticulata. So, the mechanism of sexual selection in aquatic animals is also complicated, which includes not only the single direction selection of female for male or male for female, but innersex competition. In this review, we also summarized the influence factors to sexual selection, which include the time and space of mate choice, human activity, other organism’s activity, the intensity and contrast of body color, predators, even adulterer, etc. Today, the existing sexual selection studies of aquatic animals mainly are focused on the fishes, sea urchins, and oysters, but little on the other aquatic animals. Thus, more studies are still needed in order to clarify the complicated behaviors and mechanisms of sexual selection in all kinds of hydrobios. The following problems are interesting for us to have further researches: (1) How do the sexual selection drive speciation of aquatic animals? (2) The genetic mechanism of diversity in sexual selection; (3) The relationship between nature selection and sexual selection; (4) The mechanism of female making choice; (5) The action of sexual selection in the germplasm protection. In addition, the classical theory of sexual selection is enduring new kind of trial, and new theory has been put forward in the fish, Symphodus tinca.

    • >PAPERS
    • The assessment of aquaculture loss caused by red tides

      2009, 33(4):610-616. DOI: 10.3724/SP.J.00001

      Abstract (3418) HTML (0) PDF 0.00 Byte (1904) Comment (0) Favorites

      Abstract:A new evaluation method of the aquiculture loss caused by red tides has been discussed in the present study. By analysing the structure of loss caused by red tide hazard, the aquaculture loss is divided into three types: direct aquaculture loss, indirect aquaculture loss and recovery cost. In addition, based on the different demand of timing, it was also divided into three grades: prediction assessment before hazard, tracking or monitoring assessment during hazard and onthespot surveying assessment after hazard. Finally, a practical example of assessing the loss of red tides occurring in Nanji Island, Zhejiang in June 2005, was given by red tides hazard loss assessment system established above. Its results were as follows: Prediction assessment before hazard: Classification is BR2. Early warning color is orange, and the second emergency measure was taken. Tracking or monitoring assessment during hazard: the direct economic loss is RMB 33 million. Onthespot surveying assessment after hazard: the direct economic loss is 31 million. The total economic loss is 42.9 million.

    • Construction of invasive risk assessment system for alien marine species

      2009, 33(4):617-623. DOI: 10.3724/SP.J.00001

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      Abstract:To assess the invasion risk of alien marine species, an invasive risk assessment system was constructed based on the characters of introduced marine species and on the analysis of literatures and expert consultancy. The system has 5 primary and 20 secondary indexes, and was constructed using analytic hierarchy process (AHP) method and trigonometrical fuzzy numbers for determining the weight of each index of the system. The invasive risks for 7 typical alien marine species were assessed, subsequently. The results of the system identified the risk grade of cordgrass Spartina alterniflora and White Spot Baculovirus as very high, Mytilopsis sallei, Karenia mikimotoi and Perkinsus spp., as high, Tilapia spp., as medium, and Scophthalmus maximus, as low levels of risk grade. Because traditional “1-9 scale” method is unable to reflect the real thought accurately, which leads to incorrect judgments, an improved scale method based on trigonometrical fuzzy numbers was used in the present assessment system, and the relative weights of both primary and secondary indexes were designed. Thus the assessing results reflect the real thought more accurately. It is believed that the construction of this risk assessment system will provide beneficial suggestions in decision makings for the managements of tackling alien marine species.

    • Studies on feed conversion ratio trait of common carp (Cyprinus carpio L.) using EST-SSR marker

      2009, 33(4):624-631. DOI: 10.3724/SP.J.00001

      Abstract (3518) HTML (0) PDF 0.00 Byte (1930) Comment (0) Favorites

      Abstract:Feed conversion ratio is very important economic traits. The improvement of it can not only save the feed, reduce the cost, but can also improve the ecomonic efficiency. There were a great deal of studies relating to feed conversion ratio of QTL in the field of farming. But few were reported on fish, especially on common carp. The linked markers and genes from the analysis of QTL are very important tools for fish breeding. Seventy ESTSSR markers were selected to analyze the genomic DNA of ninetytwo progenies derived from the common carp (Cyprinus carpio L.) fullsib family, which come from the cross between progenies of Barbless carp and Hebaocold tolerance red carp. 70 makers segregate according to Mendel's Law, of which 48 segregate in 1∶〖KG-*2〗1 model and other 22 in 3∶1 model. Ma kers of “1∶1” segregation model were used to made marker regression analysis based on pseudotestcross strategy using Windows Map Manager 2.0. Six loci were discovered that had impact on feed conversion efficiency (HLJE335,HLJE25,HLE547,HLJE92,HLJE203,HLJE231) and two of them had a significant impact on this trait. ESTSSR markers were used to screen the nucleotide and protein database in the National Center for Biotechnology Information (NCBI) in order to find the homologies. High sequence homologies of HLJE547 marker were observed with the nucleotide sequence of ATPase, H+ transporting, lysosomal gene of zebra fish and protein sequence of zebra fish ATPase, H+ transporting, lysosomal protein. Homologous were 74%, 94% respectively. ATPase, H+ transporting, lysosomal protein is a component of vacuolar ATPase (V-ATPase), which were detected in iontransporting epithelium of the a blowfly rectum. In addition, there were a great deal of lysosome in epithelium of the Tachypleus tridentatus Leach rectum. Fish as the first key link of craniota may have some similar function with invertebrate. This indicates that V-ATPase may affect the function of digestive system through affecting the H+ transporting on lysosomal, which needs to be proved for further study.

    • The nutrition of fatty acid and amino acid analysis of four genealogies Pseudosciaena crocea (Richardson)

      2009, 33(4):632-638. DOI: 10.3724/SP.J.00001

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      Abstract:This experiment was conducted to analyze general nutrional components, the amino acid and fatty acid composition of dorsal muscles of four Pseudosciaena crocea genealogies such as Daiquyang, Guanjingyang, their positive cross, and negative cross genealogies.It showed that there was no significant difference(P>0.05) of protein and ash among the four genealogies.The fat content of negative cross genealogy had significant differences compared with Daiquyang,Guanjingyang and the positive cross genealogies.The Guanjingyang genealogy had the highest fat content(13.51%),while the negative cross genealogy had the lowest fat content(10.59%).The water content was inversely proportional to the fat content. The water content of negative cross genealogy compared with Daiquyang,Guanjingyang and positive cross genealogy also showed significant difference.The content of total amino acid of Daiquyang,Guanjingyang and their positive cross,negative cross genealogies was 57.83%±1.19%, 54.45%±1.01%,54.80%±2.97% and 58.39%±0.71% respectively.There was significant difference of total amino acid content of Daiquyang and negative cross genealogies compared with that in Guanjingyang genealogy (P<0.05).The content of essential amino acid was 30.08%±0.52%,28.05%±0.35%,28.40%±1.76% and 29.97%±0.41% respectively.The difference of essential amino acid content of negative cross and Daiquyang genealogy compared with the Guanjingyang genealogy was significant(P<0.05).The content of tasterelated amino acid was 24.56%±0.26%, 23.45%±0.24%, 22.88%±0.61%and 25.39%±0.11% respectively,and the difference between negative cross and the other three genealogies was significant (P<0.05). In addition, the negative cross genealogy had the highest content of PUFA and DHA in the four genealogies and the difference was significant (P<0.05). The difference of EPA content between Daiquyang amd the positive cross genealogy was significant (P<0.05). These results suggested that the negative cross and Daiquyang genealogy were superior to other two genealogies based on general nutrional components, amino acid and fatty acid. It showed that the difference of nutritional components of dorsal muscle among the four genealogies may attribute to parental inheritance when they are cultured under the identical conditions. Combined with improving culture environment and nutritional compositions of diet,the quality improvement of Pseudosciaena crocea could be realized by genealogy selection.

    • Gynogenesis induced by heterogenous sperms in summer flounder Paralichthys dentatus

      2009, 33(4):533-541. DOI: 10.3724/SP.J.00001

      Abstract (3898) HTML (0) PDF 0.00 Byte (2012) Comment (0) Favorites

      Abstract:Effective methods for induction of diploid gynogenesis of the summer flounder Paralichthys dentatus are needed to initiate monosex culture, which will allow farmers to take advantage of the more rapid growth and larger size attained by females and will maximize profit potential for the culture of this species.To induce diploid gynogenesis in summer flounder, eggs were fertilized with heterogenous cryopreservative sperm of Lateolabrax japonicas, which was irradiated with UV ray to be genetically inactivated, and cold shock was used to prevent extrusion of the second polar body. Results indicated that, just like the homologous sperm of summer flounder, the cryopreservative sperm of Lateolabrax japonicas could induce diploid gynogenesis of summer flounder. Diploid gynogenesis was induced by activating egg development with UV irradiated sperm (80 mJ/cm2, the homologous or the heterogenous sperm) for 5 min in seawater at 18 ℃, and then subjecting the eggs to cold shock in 3 ℃ seawater for 45 min. And the dipoid gynogenesis induction rates of the homologous sperm and the heterogenous sperm were 32.66%±7.03% and 28.00%±6.48%, respectively. It was certified that the larvae were gynogenetic diploid by morphology, flow cytometry and microsatellite DNA analysis. It is the first time to report effective methods for induction diploid gynogenesis of the summer flounder by heterogenous cryopreservative sperm of Lateolabrax japonicas, which provides technological measures for sex control and genetic improvement of summer flounder.

    • Effects of dietary n-3 HUFA on growth performance and lipid metabolism in juvenile black sea bream, Sparus macrocephalus

      2009, 33(4):639-649. DOI: 10.3724/SP.J.00001

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      Abstract:Six diets containing graded levels (0.79,0.83,0.85,0.88,0.92 and 0.94%) of n-3 highly unsaturated fatty acids (n-3 HUFA) with a constant DHA/EPA, were fed randomly to six groups (Diet 1-6) of fish with triplicate [average weight, (8.08±0.09) g; 20 fish/replicate] to determine the effects of dietary n-3 HUFA on growth and lipid metabolism in juvenile black sea bream (Sparus macrocephalus). Results of the 8week growth trial showed that: (1) Hepotasomatic index (HSI) and intraperitoneal fat (IPF) ratio of juvenile black sea bream decreased with the increase of dietary n-3 HUFA and the values in Diet 5 and 6 were significantly lower than those in other four groups (P<0.05). Adipocyte diameter in IPF was decreased by dietary n-3 HUFA and the significance occurred between Diet 3 and 6. Lipid content in muscle was significantly affected by dietary n-3 HUFA and reached the bottom in Diet 4. No significance was found in whole body proximate composition of juvenile black sea bream. Concerning the fatty acid composition, ∑SFA and the main constituent,C16:0 were negatively correlated with dietary n-3 HUFA, while ∑n-3 HUFA content was obviously increased among all the treatments. DHA to EPA ratios (DHA/EPA) in liver, dorsal muscle and IPF were unchanged with the supplementation of dietary n-3 HUFA. Quadratic analysis based on weight gain rate (WGR) indicated that dietary n-3 HUFA requirement for black sea bream was 0.87% DM. (2) Determination of fatty acid synthase (FAS) and hormonesensitive lipase (HSL) activities showed that FAS activity kept constant in Diet 1 to 4, and then decreased significantly with further increment of dietary n-3 HUFA. HSL activity in IPF increased significantly with the elevation of dietary n-3 HUFA level and increased to almost two times in Diet 6 (0.94%n-3 HUFA). Gene expression of FAS changed following the similar trend with FAS activity in liver; while expression of HSL in IPF was positively correlated with dietary n-3 HUFA level. In summary, we can deduce that n-3 HUFA regulation in lipid metabolism was realized by affecting the lipogenesis and lipolysis at the same time.

    • Microsatellite analysis of gynogenetic diploids induced by heterologous sperm in Chlamys farreri

      2009, 33(4):542-548. DOI: 10.3724/SP.J.00001

      Abstract (3568) HTML (0) PDF 0.00 Byte (1949) Comment (0) Favorites

      Abstract:The Zhikong scallop Chlamys farreri plays an important role in the shellfish breeding in the north of China and the gynogenesis in this scallop has developed rapidly in recent years. In this study, the meiotic division of the eggs of C. farreri was activated by ultraviolet (UV)irradiated heterogenous sperms of Pacific oyster Crassostrea gigas and the gynogenetic diploids were induced by blocking extrusion of the second polar body from fertilized eggs with 6dimethylaminopurine (6-DMAP). To assess homogeneity and isogenedity between the progeny and their parents, microsatellite technique were used. Then 3 pairs of common microsatellite primers to be used for genetic analysis of offspring were screened from 21 pairs of microsatellite primers among which 11 pairs were indentified in C. farreri and 10 pairs in C. gigas by optimizing the condition of polymerase chain reaction(PCR). Under the optimal condition, PCR was performed for analyzing the genetic character of the gynogenetic offspring. It was found that the 3 loci could amplify clear fragments in these offspring. The results of genetic analysis showed that the gene of offspring without gene contamination of male parent C. gigas was completely inherited from female parent C. farreri. Microsatellite locus and centromere recombined at the 3 polymorphic loci and the recombination rates were 40%, 55%, and 35%, respectively. This study confirmed that it was feasible to achieve diploid gynogenetic progeny of C. farreri by heterologous sperm’s inducement while it was difficult to get an absolute homozygotic population once in manner of suppression of the meiosisⅡ, because there was high heterozygosity in the first meiogynogenetic generation owing to crossover between the homologous chromosome during the meiosisI.

    • Genetic structure analysis of wild and cultured populations of the Japanese sea urchin (Strongylocentrotus intermedius)using microsatellites

      2009, 33(4):549-556. DOI: 10.3724/SP.J.00001

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      Abstract:In this research,we have analyzed the genetic diversity of wild and cultured populations of 28 polymorphic microsatellite markers for Japanese sea urchin (Strongylocentrotus intermedius) via mining EST database of relative species (S. purpuratus). S.intermedius belongs to Echinodermata, Eleutheroze and Echinodea, which is distributed in the northeastern Japan, the Korean Peninsula and the Far East coast of Russia. It was introduced into China by Dalian Fisheries University in 1989 and has been breeding for more than 6 generations. ESTSSRs recently have been commomly used in genetic investigations. The flanking sequence of ESTSSRs is so highly conserved that the primers can be tansferred between similar species. Therefore, in this experiment, the primers were designed with the EST sequence of S. purpuratus from the database in GenBank (http://www.ncbi.nlm.nih.gov/dbEST/index.html), then polymerase chain reaction (PCR) was used to identify the effective primers that matched with the S.intermedius which was related to S. purpuratus. Results showed: A total of 91 different alleles were found and the number of alleles in each locus ranged from 2 to 6.In the 2 populations,the average number of alleles was both 3.071 4; the number of mean valid alleles was 2.231 9 and 2.227 1 respectively; the value of average observed and expected heterozygosity were 0.523 4,0.536 5 and 0.486 8,0.499 3 respectively; the mean PIC was 0.447 7 and 0.439 6. The difference in genetic diversity among the 2 populations was not significant. The genetic simi1arity coefficient of the 2 populations was 0.758 7,and the genetic distance of the populations was 0.276 2. Chisquare test was used to analyze the genotypes based on HardyWeinberg equilibrium,the P value showing that 50% loci departed from HardyWeinberg equilibrium. The results of the Ftest suggested that both the two populations have surplus heterozygous, and there is no obvious inbreeding phenomenan in the two populations. The results also displayed low genetic differentiations between the 2 populations,and the variance was mainly due to individual difference among the populations. Although the cultured population has been breeding for several generations, there is no significant difference on genetic polymorphism with the wild population.

    • Optimal dietary animal to plant protein ratio for the pacific white shrimp, Litopenaeus vannamei at two salinities

      2009, 33(4):650-657. DOI: 123456

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      Abstract:Protein is one of the most important components in crustacean diets, which should be optimally utilized to achieve faster growth rate of the cultured animals. To enable the preparation of a costeffective feed at the farm level, the ratios of protein derived from the various animal and plant sources in the feed need to be optimized. In this study, the factorial effects of ambient salinity and animal to plant protein ratio on growth and survival of juvenile white shrimp, Litopeneaus vannamei were investigated. Six diets were formulated using soy protein concentrated and fish meal as plant and animal protein sources respectively, of which the dietary animal to plant protein ratios were 0∶38, 7∶30, 14∶23, 21∶16, 29∶8 and 36∶0 approximately. Each diet was fed to three replicate groups of shrimp for 40 d. The results showed that dietary animal to plant protein had significant effects on the shrimp weight gain, hepatosomatic index (HSI), conditional factor, survival rate and the hepatopancreas soluble protein. Weight gain increased when the dietary animal to plant protein ratio increased, while for other indexes measured, they exhibited the tendency of increased first with the dietary animal to plant protein ratio increasing, and then decreased slightly. Increasing salinity significantly increased the weight gain, survival and the conditional factor, and significantly decreased shrimp HSI, while no significant differences of salinity were observed in shrimp HSI. Shrimp weight gain, survival and HSI were significantly affected by the interaction between ambient salinity and dietary animal to plant protein ratio, while shrimp conditional factor and the hepatopancreas were not significantly affected by the interaction between ambient salinity and dietary animal to plant protein ratio. Combined, the results in this study indicate that dietary amino acids content and profile would change with the change of dietary animal to plant protein ratio, and though L. vannamei has different demand for protein, it could be met by regulating dietary animal to plant protein ratio in the practical diets. The Brokenline model analysis showed that the optimal dietary animal to plant protein content in the diets for the white shrimps at the salinity of 3.0 and 22.0 ranged from 29.12∶7.79 to 30.29∶6.71 and from 26.05∶10.95 to 29.03∶7.437 approximately.

    • Genetic variation analysis on the first generation from matings and crosses of two geographic populations in Meretrix meretrix

      2009, 33(4):557-564. DOI: 10.3724/SP.J.00001

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      Abstract:The fluorescent amplified fragment length polymorphism (fAFLP) technique was applied to analyze the genetic variation of 4 F1 groups (SS,JJ,SJ,JS) produced from matings and crosses of Shandong stock and Jiangsu stock in Meretrix meretrix. A total of 255 AFLP loci amplified with three pairs of primers were obtained from 96 individuals. The percentage of polymorphic loci ranged from 91.98% to 96.02% of 4 groups and the overall polymorphic loci percentage reached 97.65%. The results of Nei’s gene diversity and Shannon’s information index showed that the level of genetic diversity from high to low was SJ> SS> JS>JJ and the degree of variation of two hybrid groups was greater than that of two mating groups. The analysis of fAFLP loci indicated the frequency of dominant gene of SJ and JS increased and the genetic structure of two hybrid groups was similar to their maternal parent’s. By analyzing genetic similarity between groups, it was found that the genetic similarity was 0.976 1 between JS and JJ, 0.945 0 between JS and SS. And the genetic similarity was 0.950 2 between SJ and SS, and 0.925 4 between SJ and JJ. It is obvious that the genetic similarity (0.907 5) between SJ and JS was the lowest. Phylogenetic tree based on the genetic distance of 4 groups with NJ method and UPGMA method showed that JS and JJ were clustered together firstly because of the nearest genetic relationship, and SJ and SS were also clustered together. But it is evident that the phylogenetic relationship between SJ and other groups was more distant. In conclusion, hybridization between different geographical populations in Meretrix meretrix is a dominant approach to increasing genetic diversity and variation degree; two hybrid groups in genetic structure were similar to their maternal parent; high level of genetic diversity and variation degree of SJ group perhaps have a direct relationship with its heterosis in growth.

    • Effects of partial replacement of fish meal by corn glutenmeal on daily ration, growth, and nutrient ingredients in muscles of Litopenaeus vannamei

      2009, 33(4):658-665. DOI: 10.3724/SP.J.00001

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      Abstract:Increasing economical concerns regarding the use of fish meal in diets for shrimp have led to the development of replacement strategies where plant protein sources have received ample attention. Corn gluten meal, being one of the available plant protein sources, which is obtained as a byproduct from corn, has a high protein level, and it is low in fiber, rich in vitamins B and E and is known to contain no antinutritional factors. In this study, the corn gluten meal as a dietary protein for juvenile Litopenaeus vannamei was evaluated. Four kinds of isocaloric, isonitrogenous diets were formulated. The control diet used fish meal, soybean meal, meat meal and peanut meal as the protein sources, and other three diets contained 5%, 10% and 15% of corn gluten meal respectively (replacing 8.6%, 17.2% and 25.8% of fish meal respectively). The juvenile shrimps with initial weight (0.013 6±0.001 0) g were fed in aquarium indoor for 45 d at 28 ℃. Effects of partial replacement of fish meal by corn gluten meal on growth, daily ration and nutrient ingredients in muscles of Litopenaeus vannamei were determined. The results showed that since dietary corn gluten meal level up to 10% did not significantly affect the mean daily ration(P>0.05), and 5% treatment had the best growth parameters. Its weight gain (WG), length gain (LG) and feed conversion ratio (FCR) was 2 542.8%, 155.1% and 1.58 respectively. WG was significantly higher than that of the control(P<0.05), LG and FCR were not significantly different from those of the control(P>0.05). The growth parameters of 10% treatment were not significantly different from those of the control(P>0.05), but those of 15% treatment were significantly lower than those of the control(P<0.05). There were no significant differences in content of moisture, crude protein, crude lipid and crude ash in muscles of Litopenaeus vannamei among treatments(P>0.05), and the total amino acids (TAA), essential amino acids (TEAA), delicious amino acids(DAA) and essential amino acids index (EAAI) in muscles of Litopenaeus vannamei in corn gluten meal treatments were also not significantly different from those of the control(P>0.05). 10% corn gluten meal in the feed for tested shrimps was suggested.

    • Molecular identification of Chlorella strains based on sequence analysisof nuclear rDNA ITS and chloroplast rbcL gene

      2009, 33(4):565-571. DOI: 10.3724/SP.J.00001

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      Abstract:Chlorella is one of the most important economically unicellular green algae, but its hundreds of varieties make it difficult to be distinguished based only on the morphological, physiological and biochemical features. To analyze the interspecies and intraspecies relationships and genetic variation of Chlorella species, two kinds of molecular markers were applied. One was the internal transcribed spacer (ITS) of nuclear rDNA and the other was the chloroplast rbcL gene encoding ribulose1,5bisphosphate carboxylase/oxygenase large subunit. The 6 〖WTBX〗Chlorella〖WTB1〗 strains studied included C. pyrenoidosa F5, F9, 820, C. protothecoides F2, C. vulgaris Cvq and C. ellipsoidea Ce. In addition to sequences retrieved from GenBank, a total 15 ITS and 15 rbcL sequences were carried out by sequence comparison, genetic distance calculation and phylogenetic tree construction, respectively. Results showed that the length of ITS region was almost the same among species intrasections, but the length of ITS1 and ITS2 varied significantly between species. However, the length of rbcL sequence was always conservative at inter and intraspecies levels. The genetic distances of 15 ITS sequences ranged from 0.000 to 0.663, and those of 15 rbcL sequences varied from 0.000 to 0.216. The phylogenetic relationships among the 6 Chlorella strains drawn by ITS and rbcL were as follows: First, C. protothecoides F2 showed very close relationship with C. pyrenoidosa F5 and F9. Second, C. pyrenoidosa 820 clustered closely with C. vulgaris Cvq. Third, C. ellipsoidea Ce showed the farthest distance with the other five Chlorella strains. This paper indicated that phylogenetic analysis and molecular identification of the genus Chlorella could be validated based on the combination of the highly variable ITS sequence and the relatively conservative chloroplast rbcL gene.

    • Lipid components of Ostrea rivularis, Paphia undulata and Pinctada martensii

      2009, 33(4):666-671. DOI: 10.3724/SP.J.00001

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      Abstract:Lipid composition of three seashells was analyzed in order to provide the basic data for the high value processing and development of seashells. The total lipids were extracted from Ostrea rivularis, Paphia undulata and Pinctada martensii with methanol and chloroform, the content of cholesterol and phospholipids was analyzed by colourimetry, the neutral lipid and polar lipid were separated by liquidliquid extraction, and the composition and content of fatty acids were analyzed by GCMS. The results are as follows∶the content of crude protein from three seashells is about 10%, the content of crude fat is about 1%, and the content of ash is above 1.3%. The content of cholesterol in crude fat is in the range of 0.07-0.12 mg/g, much lower than 3.55 mg/g of eggs, much lower than 5.40 mg/g of squid, therefore three seashells for human consumption do not involve the danger of high cholesterol intake. The content of phospholipids from three seashells is in the range of 14%-34%, much higher than 6.67% of soybean, but lower than 45.2% of yolk, in addition, phospholipids of seashell are also rich in EPA and DHA, therefore seashell phospholipids have a high nutritional value. The content of polar lipid in crude fat is in the range of 27%-45%, the content of neutral lipid is in the range of 51%-62%, and polar lipid composition is mainly phospholipids. The 30 kinds of fatty acids are detected from seashells lipid, the characteristics of fatty acid composition are similar in total lipid, neutral lipid and polar lipid, and the total content of saturated fatty acid is in the range of 32%-44%, mainly C14∶0, C16∶0 and C18∶0; the total content of monounsaturated fatty acid is the range of 9%-17%, mainly C16∶1, C18∶1 and C20∶1; the total content of polyunsaturated fatty acid is in the range of 41%-50%, mainly C20∶3, C20∶5(EPA), C22∶2 and C22∶5(DHA), especially the content of EPA and DHA is in the range of 15%-28%. The content of EPA in Ostrea rivularis and Paphia undulata is higher than that of DHA; the content of DHA in Pinctada martensi is higher than that of EPA. Therefore, Pinctada martensii is suitable for developing the functional food of healthy brain and prevention of Alzheimer′s, and Ostrea rivularis and Paphia undulata are suitable for developing the functional food of prevention of cardiovascular disease.

    • The action mechanism of exogenous gonadotropin inducing spermatogenesis and maturity of testis in Japanese eel, Anguilla japonica

      2009, 33(4):572-580. DOI: 10.3724/SP.J.00001

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      Abstract:Immunohistochemical localization of LHR and AR was performed by using rabbit antiserums against leuteinizing hormone receptor (LHR or choriogonadotropin receptor CGR) and androgen receptor (AR). The aim of present study was to reveal the endocrine mechanism of exogenous gonadotropin (CPH and hCG) inducing spermatogenesis in testis of Japanese eel. The results showed that the testis and spermatogenesis in Japanese eel displayed very marked changes after hormone injection. The observation of histological section showed the testis of eel was at the stage of spermatogonia proliferation before hormone treatment, while ten days after the injection of the combination of two gonadotropins, the spermatogonia mitosis and number increase of primary and second spermatocytes in the lobule of testis of male eel were seen. Thirtyfive days after hormone treatment, a lot of primary and second spermatocytes in the lobule of testis as well as some sperms in the lumen of seminiferous tubule, except a few spermatogonia near reproductive epithelium, could be seen. Eightthree days after hormone treatment, the spermatogenesis and the development and maturity as well as spermiation of testis in male eel completed. Immunohistochemical staining results further revealed that LH or CG receptors immunoreactivity was located on the reproductive epithelium showing strong immunopositive reaction before hormone treatment, while LH receptors immunoreactivity was located on the cytomembrane of Sertoli cells, Leydig cell or interstitial cell, spermatogonia, primary and second spermatocytes showing strong immunopositive reaction after hormone treatment. Before hormone treatment, androgen receptors were located on the cytomembrane of spermatogenic cells and reproductive epithelium, while AR were located within nucleus or in cytoplasm of these spermatogenic cells, but spermatids and sperm showed immunonegative reaction. The results demonstrated for the first time that the action mechanism of two hormones inducing spermatogenesis and maturity in eel is mediated by their corresponding receptors. Thus, the present study will provide scientific basis for the dosage and intervallic time of injecting hormones during artificial propagation of eel.

    • Studies on elimination regularity of doxycycline residues in Ictalurus punctatus at different temperatures

      2009, 33(4):672-678. DOI: 10.3724/SP.J.00001

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      Abstract:Doxycycline (DOTC) is a tetracycline broadspectrum antibiotic widely used for the treatment of systemic bacterial infections. The broadspectrum antibiotic doxycycline (DOTC) has commonly been used in channel catfish aquaculture to prevent and treat bacterial diseases. In order to study the tissue distribution and elimination of doxycycline (DOTC) in channel catfish (Ictalurus punctatus), continuous (for 5 days) oral administration(20 mg/kg body weight) of DOTC has been investigated in healthy channel catfish at (18±1) ℃ and (28±1) ℃water temperature. This is first report on tissue distribution and residues of DOTC in channel catfish. After repeated oral administration for 5 consecutive days, channel catfish (Ictalurus punctatus) were killed and the muscle and skin, liver, kidney tissues were collected after 1,3,5,7,9,12,15,24,30,40 d of oral administration. Doxycycline (DOTC) in tissues was determined by using high performance liquid chromatography (HPLC) with UV detection. The mobile phase was 0.01 mol/L oxalic acid∶acetonitrile∶methanol (70∶20∶10, V/V/V) and UV detection set at 350 nm. Analyte was extracted with Mcllvaine bufferEDTA extraction, whose pH was 4.0. DOTC was eluted from Oasis HLB column with 2 mL methanol. The standard curves were linear in the investigated ranges, 0.05-10.0 μg/g of DOTC in tissues. The correlation coefficient for DOTC was both r≥0.9964.The recovery of DOTC was from 75.13%to 92.05%.The withinday and daytoday precision expressed by CV was less than 10% at two drug levels (0.1 and 1.0 μg/g).The detection limit for DOTC in muscle and skin, liver, kidney was 10,20 and 20 μg/kg respectively. The results show that the depletion speed of DOTC in channel catfish closely relates with the water temperature. The elimination rate was markedly different at different water temperature in the same tissues. The depletion of DOTC residue in channel catfish was faster at high water temperature. To compare with other tissues, the elimination of DOTC residue in muscle and skin was other slowest, liver was the main reservoir of DOTC residue in channel catfish. The speed from fast to slow is muscle and skin,kidney,liver. It took liver as target tissue in this experiment. According to the maximum residue limit(MRL) of 300 μg/kg in liver, the withdrawl period should not be less than 30 d and 55 d respectively at (18±1) ℃ and (28 ± 1) ℃water temperature. This study aims to provide a theoretical basis to make dosage regimen and withdrawl time at different water temperatures.

    • A rapid immunochromatography test format for Aeromonas caviae detection

      2009, 33(4):679-684. DOI: 10.3724/SP.J.00001

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      Abstract:Aeromonas caviae is a pathogenic bacterium that can infect various farmed fishes, and cause high mortality. Thus, a rapid and sensitive technique is necessary for the detection of the bacteria to prevent further economic losses. For rapid and simple examination, an immunochromatographic lateral flow assay system (GICA) was developed. In GICA, the rabbit antiA. caviae antibody applied to nitrocellose membrane acts as a capture reagent for the target analyte in a sample, and the monoclonal antibody against A. caviae conjugated with colloidal gold acts as signal generator. Sodium citrate was used as reducer to donate electrons to the positively charged gold ions in solution and colloidal gold particles was produced. The size of colloidal gold particles was checked by a transmission electron microscope (TEM), and the TEM images showed the average diameter of colloidal gold particles was almost the same size: approximately 20.0 nm in diameter. We produced monoclonal antibody against A. cavia by hybridoma technology. After cloning, three strains of hybridoma named 0E10,1C4 and 1F10 were obtained. An absorbing pad, often paper, is attached to notrocellose membrane. On the nitrocellose membrane,rabbit anti A. caviae antibody was used as a capture antibody at the test line (T) and goat antimouse IgG antibody was used as the capture antibody at the control line (C). A conjugate pad which was attached to the notrocellose membrane contains gold particles conjugated with 1F10 monoclonal antibody specific to the analyte being detected. A sample pad, usually glass fiber, is attached to the conjugate pad. During detection, the liquid sample migrates by capillary diffusion through the conjugate pad, rehydrating the gold conjugate and allowing the interaction of the sample analyte with the conjugate. The complex of gold conjugate and analyte then moves onto the notrocellose membrane and migrates towards the test line (T), where it becomes immobilized and produces a distinct signal in the form of a sharp red line. A second line, a control line(C), was also formed on the membrane by excess gold conjugate, indicating the test is complete. The preliminary feasibility study of this method was described as follows:the sensitivity of the GICA test strip towards A. cavia was high with a detection limit of 1×106 CFU/mL, The specificity of the assay is 100% with no crossreaction being observed with thirteen bacteria of aquaculture pathogens such as A. hydrophila, A. sobria, Vibrio alginolyticus etc., Accurate reading time needed for confirmation of the assay can be completed in 5 min with a liquid sample of 100 μL, The GICA test strip is stable at room temperature for 6 months or more (data not shown). This study indicated that the GICA test strip assay system provided high sensitivity and specificity for the detection of A. cavia at low concentration. The assay is rapid, simple, cheap, and does not require any sophisticated equipment. Thus, the GICA test strip will be a useful technique for rapid diagnosis of A. cavia infection.

    • Plasmic biochemistry indicators, classification of blood cells and haecytogenesis of wild Euchiloglanis davidi

      2009, 33(4):581-589. DOI: 10.3724/SP.J.00001

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      Abstract:40 wild Euchiloglanis davidi were studied on the plasmic biochemistry indicators, classification of blood cells and haecytogenesis. In the plasmic biochemistry indicators, there were (48.35±3.60) g/L total protein, (8.73±0.54) g/L albumin, (39.63±3.10) g/L globubin, (5.05±2.14) mmol/L triglyceride, (16.66±3.64) mmol/L cholesterol, (1.01±0.43) mmol/L very low density lipoprotein (VLDL), (5.27±2.43) mmol/L glucose in plasma. These indexes indicated that albumin, globubin, triglyceride and cholesterol were positively high, but glucose negatively low in plasma of freshwater fish. These characteristics showed that E. davidi has large activity amount and is coldwater fish, which utilizes lipid and protein as main energy resource. In corpuscle of the fish, there were erythrocytes, ghost cells, lymphocytes, thrombocytes, neutrophilic granulocytes and monocytes to be found. By counts with blood smears, there were 96.58% erythrocytes (therein, 65.92% erythrocyte and 30.66% erythroblast)and 3.42%leucocytes (therein, 2.37% lymphocytes, 0.85% thrombocytes, 0.14%, neutrophils and 0.07% monocytes). Most original corpuscles were found in pronephros and mesenephros and some original leucocytes were also found in spleen, but no original corpuscles were found in liver and blood. In E. david, pronephros and mesenephros are the principal hematopoietic organs. Erythrocytes mainly come from pronephros and leucocytes mainly come from mesenephros. Some leucocytes are also produced in spleen.

    • Studies on preparation and property of the edible walleye pollock (Theragra chalcogramma) skin gelatin/chitosan composite films

      2009, 33(4):685-691. DOI: 10.3724/SP.J.00001

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      Abstract:The properties of skin gelatin films of walleye pollock (Theragra chalcogramma) were examined before chitosan was added to improve the properties of the fish gelatin films. The interaction and compatibility of various composition in the composite film were also researched, which would provide theoretic basis and scientific gist for the development of edible films in food industry. Skin gelatin of walleye pollock had good filmogenic property, while the property of composite film was improved greatly with the addition of chitosan. Fish gelatin was miscible with chitosan and the gelatin/chitosan composite films had excellent optical transmittance (higher than 88%). The mechanical property of fish gelatin film was lower, which was examined by texture analyzer. The addition of 80% chitosan and 60% chitosan increased the tensible strength and elongation at break to 182.48% and 430% of fish gelatin. Differential scanning calorimetry (DSC) analysis showed that the thermal denaturation temperature of gelatin/chitosan composite films was 10℃ higher than that of fish gelatin film. Chitosan modified fish gelatin film by decreasing water solubility and WVP sharply, but made hygroscopicity slightly higher. The results suggested that fish gelatin/chitosan composite films exhibited better mechanical property, thermal stability and water resistance than fish gelatin film. Fourier transform infrared spectroscopy (FTIR), Xray diffraction (X-RD) and Scanning electron microscopy (SEM) analysis revealed that chitosan effectively interacted with fish gelatin to form a new macromolecule, and fish gelatin/chitosan had good filmogenic property.

    • Isolation and preparations of monoclonal antibodies against lectin from mud crab, Scylla serrata

      2009, 33(4):590-596. DOI: 10.3724/SP.J.00001

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      Abstract:Lectin plays an important role in nonspecific immunity of invertebrates by including bacterial agglutination, nonselfrecognition, as opsonin and enhancing cell phagocytosis. In this paper, the author reported the purification and monoclonal antibodies (mAbs) preparation of lectin from mud crab, Scylla serrata, as well as the application of mAbs for lectin location by indirect immunofluorescence. Lectin was isolated from hemolymph of mud crab by affinity chromatography using Sepharose 4B coupled with NAcetylDglucosamine (GlcNac) which could specifically bind to mud crab lectin. The serum of S. serrata was applied to the column and the lectin binding to the column could be eluted by 0.45 mol/L NaCl TrisHCl buffer with a single peak. The agglutinating specific activity of elution peak was equated 32.6 times of original serum, and SDSPAGE showed the two main components of lectin with 87 ku and 79 ku. Seven hybridoma cell lines named BF82 etc., secreting antibodies against mud crab lectin, were prepared by hybridoma technique. Ascitic fluids of mAbs were 1∶104-1∶105 titers of indirectELISA by coated purified lectin. The mAbs could inhibit the hemagglutination of mud crab lectin with 1∶32-1∶64 titers. The Ig isotype for all seven mAbs were IgG1.The Western blots displayed that the mAbs specifically binded to the bands of 87 ku and 79 ku. Haemocytes of S. serrata were used for location of lectin by using indirect immunofluorescence technology; the results suggested that lectin is mainly distributed in the granules of granule hemocyte and the membrane of hyaline hemocyte.

    • Preliminary study on volatile flavor compounds of silver carp meat byHS-SPME coupled with GC-MS and GC-O

      2009, 33(4):692-696. DOI: 10.3724/SP.J.00001

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      Abstract:To survey silver carp (Hypophthalmichthys molitrix) meat flavor, the volatile compounds were extracted and concentrated by headspace solid phase micro extraction (HS-SPME), and evaluated by gas chromatographyolfactometry (GC-O), and were identified by gas chromatographymass spectrometry (GC-MS) in this study. GC-O was performed by a group of eight assessors using the detection frequency methods, and the odors of detected compounds were described as well. Twentynine volatile compounds were identified by GCMS, among those most of them were carbonyls and alcohols, the percentage of which is up to 88%. According to detection frequency analysis, twelve volatile compounds and four unknown compounds possessed odor activity in silver carp meat. They are characterized by fishy, grassy, metallic, oil oxidation and mushroom odors, respectively. Their synergistic action formed the characterized odor of silver carp meat.

    • Mathematical analysis of effects of morphometric attribute on body weight of largemouth bass (Micropterus salmoides)

      2009, 33(4):597-603. DOI: 10.3724/SP.J.00001

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      Abstract:The effects of morphometric attributes on body weight of largemouth bass were analyzed. Data of total length (X1),standard length(X2),body depth (X3),body width (X4),interorbital distance (X5),head length (X6),snout length (X7),caudal peduncle length (X8),caudal peduncle depth (X9) and body weight (Y)were collected from culture pond in this study. Correlation analysis, path analysis and multiple regression were used. The correlation coefficients among the morphometric attributes were calculated, then the total length,body depth,head length and caudal peduncle depth were eliminated from the variable data, because all of them were colinear with standard length. Snout length and caudal peduncle length were also kicked out from the variable data because of no significance in multiple regression equation. The three morphometric attributes (X2, X4, X5) were used as independent variables, and body weight (Y) was used as a dependent variable for path analysis. Path coefficients (Py.x), determination coefficients (dy.x) and correlation index (R2) were calculated in path analysis. The results showed that the three independent variables significantly affect body weight with correlation coefficient 0.942, 0.979, 0.928 (P<0.01 ) respectively. The path coefficients (Py.x) of the body width,standard length and interorbital distance to the body weight have all reached a level of significance. These attributes were very indicative of determining the body weight, among them the body width weighted the most (P4= 0.599) to the body weight, it was a key effective factor, and standard length and interorbital distance weighted the second and the third (P2=0.231,P5=0.189). Judged from the results of high correlation index (R2= 0.980) , the main variables (X2, X4, X5) have been selected. We have obtained the multiple regression equation to estimate the body weight as LgY=1.065 + 0.765 LgX2+ 1.441 LgX4 +0.543 LgX5. This paper provides theoretical evidence and perfect measure target for selective breeding of largemouth bass.

    • Model for separating efficiency assessment of separator devices rigged in multi level beam trawls

      2009, 33(4):697-703. DOI: 10.3724/SP.J.00001

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      Abstract:Research on species selection is one of important potential measure to alleviate the bycatch and discard problem of trawl fisheries. More and more research showed that parametric models, which have been widely used in analysis of selective devices of trawls, were not only important to assess and compare the performance but indicative to improve the construction of the selective devices. In this paper, a model for assessing the separating efficiency of separator devices rigged in multilevel beam trawl fishing offshore of the East China Sea, in which the size selection of separator panel was taken into account and encounter probability (presented by ogive, Logistic curve) of individual entering trawl was regarded as dependent upon the individual size and then presented by ogive (Logistic curve), was developed based on the principle of Select model. Fitness of model for size frequency of Parapenaeposis hardwickii and Cynoglossus abbreviatus caught by multilevel beam trawl rigging separator panel with 60 mm mesh size and the maximum likelihood estimates of parameters showed the model was applicable to assess the separating efficiency of panel. According to the discussion, the model could be applied to the other selective devices and simplified by hypothesis test method according to different situation. Additionally, we think the encounter probability of individual, which was independent of size selection of selective device, could be used to analyse the responding behavior of aquatic animal.

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