YAN Xiwu , 张跃环 , HUO Zhong-ming , SUN Huan-qiang , YANG Feng , ZHANG Guo-fan
2010, 34(6):701-709. DOI: 10.3724/SP.J.1231.2010.06288
Abstract:The study on phenotypic traits of different shell color strains F1 and F2 (C:the control strains; Tr; twoband red strain; Tw:twoband white strain; Ab:agateblack strain; Or:oceanred strain; Pw:pearlwhite strain; Z:zebra strain; W:wave strain.)of Manila clam was conducted in September 2007.The results showed that the shell length of Pw F1 was the greatet,and significantly greater than others except Tr (P<0.05),the shell length of Z was the smallest,and significantly smaller than others(P<0.05),there were no obvious differences in shell length among C,Or,W,between Tr and Tw,among Tw,Ab,Or,among Ab,Or,W(P<0.05).Both fresh weight and fecundity of Pw F1 were the greatet,that of Tr were the second greatet and significantly greater than others(P<0.05),those of Z were the smallest,and significantly smaller than others(P<0.05),there were no remarkable differences between C and W,among Tw,Ab,Or in fresh weight and between C and Z,among Tr,Ab,Or,W in fecundity(P<0.05).No significant differences in eggdiameter,the fertilized rates and size of D larvae were observed among all strains(P<0.05),the hatching rate of C was significantly lower than those of other strains except Tw(P<0.05).By end of pelagic period(9 d),the shell lengths of W and Z were significantly greater than those of other strains(P<0.05),there were no remarkable differences in shell length among Tw,Ab,Or,Pw,Z,W(P>0.05); the sequences of survival rates were following:Ab,Z>Or,W>Tw,Pw>Tr>C,there were significant differences among them(P<0.05).No remarkable difference in both metamorphic time and metamorphic size among all strains was observed(P >0.05).There were such a sequence in metamorphic rates as follows:Ab>Z>Tw >Or>Pw>W>Tr>C,there were significant differences among them(P<0.05).By the end of indoor rearing period(240 d),the shell length of Ab were significantly greater than those of other strains except Or,Tr(P<0.05),the shell length of Z were significantly smaller than those of other strains except C,W(P<0.05),there were no remarkable differences in shell length between C and Pw,W,Tr,Tw,Or,and between Tw and Pw; the survival rate of Z was remarkably higher than those of other strains except Ab(P<0.05),the survival rate of C was remarkably lower than those of other strains except Tr,Tw(P<0.05).On day 360,both shell length and fresh weight were the greatet and remarkably greater than those of other strains(P<0.05),the shell length of Ab and Z were significantly smaller than those of other strains(P<0.05),the survival rate of Ab and Z were significantly higher than those of other strains except Or,Pw(P<0.05),the survival rate of Or was significantly higher than that of C(P<0.05),there was no remarkable difference in survival rate among other strains(P<0.05).The order of relative yields was as follows:Z(678.63%±56.80%)>Ab(554.88%±69.42%)>Or(527.23%±76.21%)>W(475.97%±90.25%)>Pw(405.90%±55.19%)>Tw(224.89%±47.85%)>Tr(178.50%±34.50%)>C(100.00%±22.10%).
陆剑锋 , WAN Quan , Ying Zhang-ming , Lin Lin , Weng Shi-bing , Ye Ying-wang , Jiang Sao-tong
2010, 34(6):801-808. DOI: 10.3724/SP.J.1231.2010.06800
Abstract:Collagen is one of the predominant proteins in the living body,and the essential protein component in connective tissue.Compared with collagen from land animals(such as pig,cow),aquatic collagen has drawn extensive attention for its excellent physiochemical properties and physiologically active functions.Chinese softshelled turtle(Pelodiscus sinensis)is a rare and precious traditional aquatic animal in China.It has a high medicinal and nutritional value with plenty of calipash collagen.The collagen from calipash of P.sinensis was extracted by pepsin digestion under acid condition.The effects of pepsin addition,substrate concentration and extraction time on yield of collagen were investigated.5 mg/mL pepsin addition,4% substrate concentration and 14 hours extraction time were established as optimal conditions for preparing calipash collagen.The extracted crude collagen was purified by salt precipitation and dialysis,and then its biochemical characterization was determined by the methods of SDS-PAGE,DSC,UV and FT-IR scanning.The result indicated that the extracted calipash collagen was of high purity and belonged to typical collagen Ⅰ with two α and one β chains.It can be assumed that the calipash collagen of P.sinensis might be a useful substitute for porcine and bovine collagens.This study laid a scientific foundation for further exploitation and utilization of the calipash collagen polypeptide of Chinese softshelled turtle.
YANG Ai-fu , ZHOU Zun-chun , SUN Da-peng , DONG Ying , JIANG Bei , WANG Xiao-yu , CHEN Zhong , GUAN Xiao-yan , WANG Bai
2010, 34(6):710-717. DOI: 10.3724/SP.J.1231.2010.06783
Abstract:Ferritin is widely distributed throughout the plant and animal kingdoms in a highly conserved conformation,which plays a key role in cellular iron metabolism including iron storage and detoxification.Vertebrate ferritin has been indirectly linked to innate immune response since their synthesis is regulated by proinflammatory cytokines.In invertebrate,ferritin has also been shown to be relevant to innate immune defense via ironwithholding strategy because it can be elevated by LPS challenge.To be better understand the sequences information and do further research on the function of ferritin,the cDNA sequence is characterized and the gene expression pattern is profiled.In this study,a fulllength ferritin cDNA sequence was obtained by constructing cDNA libraries of sea cucumber(Apostichopus japonicus),which was 792 bp long,and includes a 133 bp long 5′untranslated region(UTR),0RF encoding 173 amino acids,and a 137 bp long 3′UTR; the molecular weight predicted was 20 ku.The 5′UTR contains a highly conserved iron responsive element(IRE)sequence.The amino acids have a conserved motif for ferroxidase center typical of heavy chains of vertebrate ferritins.Sequence comparison showed that it reached the highest similarities of 84% with sea cucumber Holothuria glaberrima and shared more identity to H-chains of vertebrate ferritins than to the Lchains.It also shared 74%-34% identity with invertebrate ferritins compared with sea star,abalone,oyster,sea anemone,nematode,crayfish and drosophila.Phylogenetic analysis revealed the A.japonicus ferritin was grouped together with the invertebrate ferritin cluster.The ferritin mRNA were identified by using RT-PCR at development stages of sea cucumber including unfertilized egg,fertilized egg,cellulous stages,blastula,gastrula,early auricularia,auricularia,late auricularia,doliolaria,pentactula,juvenile and in different tissues of young sea cucumbers including intestines,coelomocytes,respiratory trees and body walls.Quantitative realtime PCR results showed that ferritin messages were low in unfertilized eggs through gastrula,however,a pronounced increase in transcript levels was observed from early auricularia to juvenile; The lowest expression level was found in the respiratory trees of young sea cucumbers; Moreover, ferritin mRNA showed no significant changes in expression after a LPS challenge in the four tissues of young sea cucumbers.This result suggests that ferritin expression might be regulated posttranscriptionally in sea cucumber.
LIU Hai-jing , liuyongxin , HOU Ji-lun , SUN Chao-hui , WANG Gui-xing , WANG Yu-feng , ZHANG Xiao-yan
2010, 34(6):718-724. DOI: 10.3724/SP.J.1231.2010.06830
Abstract:In this study reproduction by meiotic gynogenetic diploids and mitotic gynogenetic doubled haploids were carried out in Japanese flounder,Paralichthys olivaceus, by the same fertilized eggs from one female.The process of meiotic diploids and mitotic diploids produced involves two steps.First,the DNA of the sperm is fragmented by UV irradiation.Next,the meiotic diploids are made by prevention of the extrusion of the secondary polocyte and the haploid embryo is made diploid by inhibition of the first cell division.This is typically done by a cold and pressure shock administered around the time of the prophase of the second meiotic and first mitotic division,respectively.All the while normal diploids with Japanese flounder sperm were also produced as controls.Normal diploids and two gynogenetic diploids were analyzed by unilocus 10 microsatellites loci.The result showed the parental genotypes were found to be heterozygous at 7 loci and heterozygous at 3 loci.6 genotypes were found in normal diploid,alleles were random combination from the parents and the types enriched.Genetic similarity indexes were 0.976 6 and 0.959 5 between female and progenies and among progeny individuals,respectively,which approached that of the inbred line.Two genotypes were found to be completely homozygous in mitotic gynogenetic double haploid.Genetic similarity indexes were 0.806 2 and 0.742 5 between female and progenies and among progeny individuals.Furthermore,mitotic gynogenetic doubled haploids were fully homozygous.Meiotic gynogenetic diploids with a high degree of genetic similarity were suitable for fixing traits of female.Mitotic gynogenetic doubled haploids with high homogeneity were suitable as clonal founders.Differently from meiotic gynogenesis diploidization is based on a prekaryogamic oocyte manipulation,in mitotic gynogenesis it is actually a postkaryogamic zygotic manipulation.The resulting mitotic gynogenetic double haploids are all homozygous but not clonal because they do not necessarily share the same alleles owing to variable chromosome segregation and genetic recombination during maternal gametogenesis.Each of them,however,can be a clonal founder capable of giving rise to an isoallelic clone,if reproduced gynogenetically.Although mitotic gynogenesis can achieve homozygote according one generation,but the survival rate of process in induction,incubation and maturation is relatively low,the establishment of clone is extremely difficult.Thus,scholars have proposed to improve the genetic similarity and fix the good traits of female with successive meiosisinduced methods.In view of distinct genetic characteristics in both diploids,as long as the successful induction,it can play an important role in fixing characters of female and clone production.Both kinds of gynogenesis can become high quality breeding materials or improved strains.
Fu Jianping , Chang Mingxian , Guo Zheng , Zhu Bangke , NIE Pin
2010, 34(6):725-732. DOI: 10.3724/SP.J.1231.2010.06819
Abstract:In living organisms,ferritin plays an important role in iron storage and detoxification.The full-length cDNA sequence of a ferritin(EsFer)was obtained from the cDNA library of the Chinese mitten crab Eriocheir sinensis stimulated with lipopolysaccharide(LPS).It has 1 118 bp in length and contains a 480 bp open reading frame(ORF)encoding 159 amino acids(aa).The 5′and 3′untranslated region(UTR)is 178 bp and 461 bp,respectively.The predicted molecular weight is 18.3 ku and the theoretical isoelectric point 5.81.A transmembrane helix was found at the position of 15-37 aa.A complete ironresponsive element(IRE)was found at the 5′ untranslated region corresponding to the nucleotide sequence at the positions of the 135-162.Quantitative realtime PCR was used to assess the mRNA expression of EsFer in organs or tissues and its induced expression in haemolymph,intestine,hepatopancreas of crabs challenged with LPS.The results showed that EsFer was expressed in all tested organs or tissues of the Chinese mitten crab,including haemolymph,muscle,intestine,gills,heart,gonad and hepatopancreas.The highest expression level was observed in hepatopancreas,while the lowest in haemolymph.The expression of EsFer in haemolymph,intestine and hepatopancreas was upregulated following the challenge with LPS.
CHEN Changsheng , JI Dehua , ZHAO Guorui , Xu Yan , SHI Xiuzhou
2010, 34(6):733-740. DOI: 10.3724/SP.J.1231.2010.06778
Abstract:20 simple sequence repeat(SSR)primerpairs were used to analyze the genetic diversity of 44 germplasm materials of Porphyra haitanensis stored in the germplasm repository.A total of 81 alleles were detected,with the number of alleles ranging from 2 to 6,and the average was 4.05,and the polymorphism information content(PIC)was between 0.15 and 0.79,with the average of 0.57.Then the 16 polymorphic alleles amplified from 3 primerpairs of SSR(Phes08,Phes03 and PC13)were used to construct the DNA fingerprinting dabatase of the 44 germplasm materials of P.haitanensis.In the constructed DNA fingerprinting database,each germplasm material has its unique fingerprinting pattern and can be easily distinguished from each other.At last the DNA fingerprints were transformed into digital fingerprints which can be recognized by computer based on appear ance and disappear ance of amplified band in each allele,and developed the corresponding software of digital fingerprints recognizer to predigest the germplasm identified process.In conclusion,the fingerprinting database will provide a basis for germplasm autoidentification and digital management of germplasm repository of P.haitanensis.
郭斌 , ZHANG Bo , DAI Fang-qun , JIN Xian-shi
2010, 34(6):741-747. DOI: 10.3724/SP.J.1231.2010.06762
Abstract:Setipinna taty,is a commercially important fish distributed from the northern East China Sea to the Bohai Sea,and plays an important role in the marine ecosystems.In recent years,the catches were mainly composed of juveniles,and characterized by lower age population structure.In the present paper,the diet composition and ontogenetic variation in the feeding habits of Setipinna taty at fork length(FL)of 35-79 mm were examined by applying the KruskalWallis test,a chisquared test and the cluster analysis,according to the total of 207 samples from seasonal bottom trawl surveys in the central part of Yellow Sea in August 2008.Based on stomach contents analysis,the Setipinna taty was a generalist and more than 40 prey species were ingested.Crustaceans,including copepodas and euphausiaceas,were the most important prey groups,accounting for 94.65% of the total food by numerical percentage.At the species level,Paracalanus parvus, Calanus sinicus, Labidocera euchaeta and Euphausia pacifica were the most important prey.The relative importance of polychaetas,gastropodas and bivalvias was comparatively low.The feeding intensity,as indicated by the percentage of empty stomach(PES)and mean stomach fullness index(MSFI),varied significantly among different classes of sizes.The MSFI increased with increasing fish size,but the PES did not present the obvious trend of change.The percentage of euphausiaceas and mysidaceas increased significantly with increasing fish size,whereas the percentage of copepodas decreased.The Levin’s standardized indexes of each group are all below 0.6,which indicated that Setipinna taty has narrow dietary breadth.And the Levin’s standardized index is less and less with the increasing of fish size,and this shows that the fish’s diet becomes more and more especial.So they can make full use of environmental resources and enhance their ability to accommodate the impact of environmental fluctuation.The mean weight of prey,besides,also becomes larger and larger with the increasing of fish size.Cluster analysis revealed that there was an abrupt change in diet composition in the fish at about 45 mm FL.Copepodas dominated the diet of fish smaller than 45 mm FL,whereas euphausiaceas and mysidaceas were the dominant prey for fish greater than 45 mm FL.The result shows that the ontogenetic variation of Setipinna taty is not related with the age,but with the length.
Ma jin , LIU Bi-lin , LU Hua-jie , LI Si-liang , CAO Jie
2010, 34(6):748-754. DOI: 10.3724/SP.J.1231.2010.06706
Abstract:According to the samples of Ommastrephes bartramii collected by Chinese squid jigging vessels from July to October in 2007 in the Northwest Pacific Ocean,the ratio of statolith maximum width(MW)to total statolith length(TSL),as an indicator of shape change of statolith,was analyzed in different sexual maturitiy stages and individual sizes using the method of ANOVA and LSD.The results indicated that the statolith grew constantly in the entire life cycle of O.bartramii.MW/TSL values between sexual maturity stage Ⅰ and Ⅱ,Ⅲ,Ⅳ,Ⅴ showed significant differences,and MW/TSL values among sexual maturity stage Ⅱ,Ⅲ,Ⅳ and Ⅴ showed no significant differences.We conclude that the statoliths of O.bartramii hardly change their shape after the sexual maturity stage I.Between the four groups whose mantle length were less than 280 mm and most other groups,MW/TSL values showed significant differences,and MW/TSL values among groups whose mantle length were more than 280 mm,showed no significant differences.We conclude that the statoliths of O.bartramii hardly change their shape after mantle length of 280 mm.Mantle length of 200 to 280 mm corresponds with sexual maturity stage Ⅰ,and the period O.bartramii to with mantling length of 200 to 280 mm corresponds to the days for O.bartramii migrate into the subArctic waters.MW/TSL might serve as an valid indicator of sexual maturity and early ontogenetic shifts for the squid O.bartramii.
董晓丽 , CHEN Songlin , JI Xiangshan
2010, 34(6):649-655. DOI: 10.3724/SP.J.1231.2010.06798
Abstract:Halfsmooth tongue sole(Cynoglossus semilaevis)belongs to Pleuronectiformes,Soleidae,Cynoglossus.It contains ZWZZ sex determination system and is a newly exploited and commercially important cultured marine flatfish in China.Females grow two to three times faster than males.Thus,the development of allfemale stock would be of significant benefit for aquaculture.To develop allfemale stock of C.semilaevis,the investigation of sex determination mechanism is necessary. A Dmrt3 cDNA fragment was cloned from C.semilaevis. The fragment was 228 bp long containing the DM conserved region.NJphylogenetic tree indicated that Dmrt3 gene of C.semilaevis had a high homology with that of Fugu rubripes, Oryzias latipes and Danio rerio. Real-time PCR showed that Dmrt3 was expressed in all seven tissues and extremely higher in male gonad(P<0.01).The expression of Dmrt3 gene in gonads of methyltestosterone treated male and neo-male was higher than that of normal males(P<0.01).But no significant difference was detected between normal female and the high temperature treated neo-male. This indicates that Dmrt3 gene is not indispensable for sex reversal.The high expression of Dmrt3 in MT treated fish is a result of methyltestosterone influence.From 43(days post hatching)to 5 months old, Dmrt3 was expressed very low.At 7 months old,the expression rose up.The highest expression of Dmrt3 occurred in ninemonthold gonad tissues. After 9 months old,the expression of Dmrt3 declined.According to morphological observation,we estimate that the ninemonth old testis is in phaseⅡ of fish gonad development stages.Large amounts of spermatogenic cells exist in phaseⅡ testis and the spermatogenic cells change to primary spermatocytes in phase Ⅲ.This implied that the expressions of Dmrt3 gene go up with the increasing of spermatogenic cells. Maybe Dmrt3 gene plays an important role in the formation of spermatogenic cells.Dmrt3 was considered not to be necessary for sex reversal.
2010, 34(6):755-761. DOI: 10.3724/SP.J.1231.2010.06669
Abstract:Due to the high economic value and market demand of Hizikia fusiformis(Harvey)Okamura(Sargassaceae,Phaeophyta),the aquaculture of this alga is now being considered for larger scale production in China.Complete artificial seedling of Hizikia via sexual reproduction is essential in both conserving the natural resource and further promoting cultivation development for this economic important species.Obtaining a large number of male and female receptacles of Hizikia with identical developmental stage is the prerequisite for increasing the efficiency and effectiveness of artificial seedling.However,the developments of the male and female receptacles of Hizikia are usually not synchronized.In the present work,we tried to research the preservation methods for isolated female receptacles of Hizikia at low and ultralow temperatures,and explore their application in artificial seedling of Hizikia via sexual reproduction.Mature plants of Hizikia were collected from the low intertidal zone at Yunao Bay,Nanao Island,Shantou,China during June when Hizikia reached the peak of sexual reproduction.The female receptacles were cut from the middle part of the plants and were maintained in seawater with room temperature of about 20 ℃.Firstly the isolated receptacles were pretreated.The pretreatments procedures included election of protectants(such as seawater,DMSO,glycerol,sucrose,DMSO+glycerol+sucrose),putting receptacles in submersed or emersed states,and the freezing steps.The receptacles were then preserved at ultralow temperature(liquid N2,-196 ℃),freezing temperature(-18 ℃),or low temperature(5 ℃)respectively.We had adopted several indicators,i.e.,the photosynthetic and respiratory activities,relative viability of cells,and the capacity of gamete release,to check the reliability of all kinds of preservation methods.The results showed that the protocols of ultra-low and freezing temperatures preservation were not ideal.However,the photosynthetic and respiratory activities,relative viability of cells,and the capacity of gamete release were nearly maintained after the receptacles were preserved for 30 d at low temperature in emersed state.This supplied a simple and safe approach of receptacles preservation.We discussed the application of receptacles preservation.Low temperature preservation of isolated receptacles of Hizikia can be applied to artificial seedling.Additionally,receptacles of Hizikia can be safely transported to another site for a long time as long as the receptacles are preserved at low temperature.
王中铎 , TAN Wei , GUO Yu-Song , LIU Li , LIU Chuwu , LIU Yun
2010, 34(6):656-664. DOI: 10.3724/SP.J.1231.2010.06799
Abstract:The entire mitochondrial DNA sequence(mitogenome)of five species,Lutjanus bengalensis, Lutjanus malabaricus, Lutjanus sebae, Lutjanus kasmira and cobia(Rachycentridae:Rachycentron canadum)were determined using long PCR and primerwalking methodology with universal primers,and their GenBank Accession numbers are FJ171339,FJ416614,FJ824741,FJ824742 and NC_011219,respectively.The mitochondrial genomic sequence in snappers and cobia are similar to other vertebrate mitochondrial genomes with respect to gene order and genomic organization.Phylogenetic relationships derived from mtDNA sequences of Lutjanus russellii, Lutjanus rivulatus, Lutjanus bengalensis, L.malabaricus, L.sebae, L.kasmira, cobia and dark banded fusilier(Caesioninae:Pterocaesio tile)were incongruent with those derived from morphological taxonomic analyses.Beyond expected,high homogeneities were observed between some lutjanids and fusilier,belonging to Lutjaninae and Caesioninae.Further efforts may clarify the phylogenetic relationships among these snappers.Moreover,we investigated the performance of all thirteen mitochondrial proteincoding genes to recover the phylogenies.According to these tests,mitochondrial proteincoding genes can be roughly classified into four groups at the level of genus:of very good(ATPase6 and cox2),good(cox1 and ND2), medium(cox3, NDl, ND4, ND4L and ND5),and poor(ND3, ND6,and ATPase8).And we inform that simple length differences and rate differences between these genes cannot account for their different phylogenetic performance.We advice the careful selected genes and their combinations should be helpful for high resolved phylogenetic analysis for different level taxon.
HUANG Xuxiong , ZHOU Hongqi , HUA Xueming , YANG Zhigang , LENG Xiangjun
2010, 34(6):762-768. DOI: 10.3724/SP.J.1231.2010.06450
Abstract:In order to assay the effects of different bubbling modes on microalgae during cultivation,the growth,biochemical composition and utilization on N and P from medium for Dunaliella salina treated with three bubbling modes(shaking flask manually,continuous aeration and CO2 bubbling)respectively were investigated in this paper.The results showed that the biomass(1.62×107ind/mL)of D.salina treated with continuous aeration was significantly higher than those of the other groups after 5 days. D.salina mainly used the nitrogen stored in cell for growth and reproduction in the first 3 days.Then N in the culture medium was dramatically absorbed by D.salina cell.While the cell of D.salina could rapidly absorb P from the culture medium after inoculation and store P in the cell for the following growth and reproduction.After first three cultured days,N content in the culture medium treated with continuous aeration was significantly lower than those of the other groups.The bubbling mode did not affect the cell of D.salina to absorb P from the culture medium significantly.The contents of total lipid,crude protein and amino acid of D.salina treated with continuous aeration were significantly lower than those of the other groups.The content of PUFAs(polyunsaturated fatty acids)of the cell treated with continuous aeration was significantly higher than the other groups.It is therefore suggested that the bubbling modes affect growth as well as cell composition of the microalgae D.salina.The growth performance and fatty acid nutritional value of the cell treated with continuous aeration improve while the amino acid nutritional value of the same cell decreases.
YU Lin-yun , BAI Jun-jie , FAN Jia-jia , LI Xiao-hui , YE Xing
2010, 34(6):665-671. DOI: 10.3724/SP.J.1231.2010.06832
Abstract:Myostatin or GDF-8,a member of the transforming growth factorβ(TGF-β)surperfamily,has been demonstrated to be a negative regulator of myogenesis and inhibited myoblast proliferation during cell cycle and myogenic differentiation in animals.In present research,two SNPs sites(C-1453T and T+33C)were identified from total MSTN gene in larghmouth bass by using PCRSSCP technique and sequencing; The mutational site of C-1453 was between the regions of E8 box and Octamer(+).the mutation of T+33C was situated in the first extron of the MSTN gene,however the mutation of the first extron has not changed the encoded amino which belonged to the synonymy mutation.The three kinds of genotypes per mutation site were obtained by PCRRFLP method and the genotype frequency of the mutation was caculated in the population of 128 individuals.A general linear model was used to statistically analyze associations of MSTN gene polymorphisms with major growth traits respectively,and the results indicated that there was no significant difference(P>0.05)by all genotypes of single mutation site.Six diplotypes,with the minor allelic frequencies of above 3%,were constructed based on two SNPs in the experiment population.The means of diplotype D2 were over other diplotypes and the means of diplotype D5 were lower other diplotypes in terms of body width,body height,body length,body weight,and snout length therfore associations analysis indicated that there were significant associations between diplotype D2 and D5 in the five growth traits of largemouth bass.The study implies that diplotypes D2 and D5 of MSTN gene could be used as the moleculer genetic marker for selection breeding.
YIN Fei , SUN Peng , SHI Zhaohong , WANG Jiangang
2010, 34(6):769-774. DOI: 10.3724/SP.J.1231.2010.06823
Abstract:A 9week feeding experiment was carried out on juvenile silver pomfret Pampus argenteus[initial weight,(4.80±0.11)g] to evaluate the effects of feed composition on weight gain,feed conversion ratio(FCR),hepatic lipase and antioxidant enzyme activities using nutriology and biochemistry.Four diets were formulated and named diet 1(fresh fish meat),diet 2(fresh fish meat mixed with formulated feed),diet 3(fresh fish meat mixed with formulated feed and solen meat)and diet 4(fresh fish meat mixed with formulated feed,solen meat and copepods).Each diet was assigned to triplicate groups.The results showed that the weight gain of silver pomfret could be affected significantly by the feed composition.The weight gain of diet 1 was the lowest among the four diets,and was significantly lower than those of other three diets.The weight gain of diet 4 was the highest,and was significantly higher than those of diet 2 and diet 3(P<0.05).The FCR of diet 1 was the highest among the four diets,and was significantly higher than those of other three diets.There was no significant difference in FCR among diet 2, diet 3 and diet 4(P>0.05).Hepatic lipase and total lipase activities of diet 1 were significantly lower than those of other diets(P<0.05),but no significant difference was found in hepatic lipase,lipoprotein lipase and total lipase activities among diet 2,diet 3 and diet 4(P>0.05).There was no significant difference in superoxide dismutase(SOD)and glutathione peroxidase(GSH-PX)activities among all diets(P>0.05).However,the lowest SOD and GSHPX activities were both in diet 1 and highest were in diet 4.The catalase(CAT)activity of diet 4 was significantly higher than those of diet 1,2 and 3(P<0.05),but no significant difference was found among diet 1,2 and 3(P>0.05).In conclusion,the only use of fish meat as a feed is not beneficial to the growth of silver pomfret.Copepods could be also used in the diet to improve the growth and antioxidant ability of silver pomfret.
CAI Mingyi , LIU Xiande , WANG Zhiyong
2010, 34(6):672-678. DOI: 10.3724/SP.J.1231.2010.06756
Abstract:In order to assess the potential applications of hybrid F1 between Pseudosciaena crocea ♀ and Nibea albiflora ♂ in the genetic breeding and the basic research,the inheritance and segregation patterns of AFLP markers in the offspring of two hybrid families(HF1 and HF2)were investigated.Totally,478 and 446 fragments were detected in two pairs of parents by using 8 selective primer combinations,respectively.In the parents of HF1,the amplified fragments included 215 female parentspecific bands(FSB),165 male parentspecific bands(MSB)and 98 mutual bands(MuB).All of these fragments were transmitted to the hybrid offspring.Among these fragments,121(56.3%)FSB,115(69.7%)MSB and 93(94.9%)MuB were transmitted to all the offspring,while the other loci were segregated in the offspring.The level of polymorphism of the FSB was comparable to that in the intraspecific hybrid F1.There was no significant difference between the average frequency of the segregated FSB and that of the segregated MSB.The inheritance and segregation patterns of AFLP markers in HF2 were similar to those in HF1,except that the proportion of segregated loci in HF2 was lower than that in HF1 and 2 nonparental bands were detected in HF2.In addition,a relative high proportion of loci showing distorted segregation was found in both HF1 and HF2.These results suggested that the newly hatched hybrid fries between P.crocea ♀ and N.albiflora ♂ contained the genome of both parents,and the phenomenon of uniparental gene elimination was not obvious.The findings would serve as the basis for further development and management of the hybrids.
ZHOU Hui-hui , Ma Hongming , ZHANG Wen-bin , XU Wei , LIUFU Zhi-guo , MAI Kang-seng
2010, 34(6):775-783. DOI: 10.3724/SP.J.1231.2010.06809
Abstract:The objective of this work was to select probiotic bacterial strains for juvenile sea cucumber Apostichopus japonicus.Three strains of potential probiotics isolated from the intestines of A.japonicus were identified by 16S rDNA sequence genotyping as Bacillus GSC-1, Bacillus GSC-2 and Enterococcus GSC-3.The safety of these potential probiotics was evaluated by immersing juvenile A.japonicus in sea water with GSC-1,GSC-2,GSC-3 or Vibrio splendidus at 100,103,105 or 107 CFU/mL respectively for 7 days. Results showed that GSC-1 and GSC-3 treatments were safe under the designed concentrations based on the mortality rates. However,GSC-2 caused significantly higher mortality at 107 CFU/mL than the blank control(P<0.05).A 20day feeding trial was conducted to investigate the effects of GSC-1 and GSC-3 on the growth performance and innate immune response of juvenile A.japonicus.Feeds were algae(Sargassum thunbergii)powder supplemented with GSC-1 or GSC-3 at 0(control)and 109 CFU/g respectively.The specific growth rate and survival rate were significantly increased in treatment supplemented with GSC-1.Comparatively,survival rate and discoloration rate were remarkably improved in treatment of GSC-3(P<0.05).Moreover,GSC-1 increased total bacteria counts and decreased Vibrio spp.counts significantly(P<0.05).Phenoloxidase and lysozyme activities in body tissue of the juvenile A.japonicus were significantly higher in the group treated with GSC-1 or GSC-3 than the control(P<0.05)while acid phosphatase and superoxide dismutase activities were remarkably higher than control in GSC-1 group only(P<0.05).Both GSC-1 and GSC-3 showed positive effects on disease resistance of juvenile A.japonicus,especially GSC-3 significantly improved disease resistance to V.splendidus exposure(P<0.05).In conclusion,both Bacillus GSC-1 and Enterococcus GSC-3 show probiotic effects,therefore they can be used as probiotics in the farming of juvenile sea cucumber A.japonicus.
LI Mingyun , 胡玉珍 , MIAO Liang , CHEN Jiong , SHI Yuhong , XUE Liangyi , XU Wantu
2010, 34(6):679-684. DOI: 10.3724/SP.J.1231.2010.06750
Abstract:In this study,the growth characteristics of four Pseudosciaena crocea groups including the selfing of DD(Daiquyang♀×Daiquyang♂),the selfing of GG(Guanjingyang♀×Guanjingyang♂),their direct cross DG(Daiquyang♀ ×Guanjingyang♂),and backcross GD(Guanjingyang♀ ×Daiquyang♂)were determined.Results after 526 days of growout showed that the final mean weights were higher for the GG(330.514 g)and the GD(336.694 g)than those of the DD(278.975 g)and the DG(243.297 g).The 1yearold DD grew in a lower rate,but after the winter,the growth rate of DD accelerated significantly:the growth rate in length was higher than the other three groups while the weight gain was not significantly different from the GG and the GD.That is,the growth potential of the DD at later stage was high.The order of the fullness on 526th day was DD
HE Lin , JIANG Ming , WU Hao , YANG Li
2010, 34(6):784-791. DOI: 10.3724/SP.J.1231.2010.06814
Abstract:A method using reversed phase high performance liquid chromatography with UV detection has been developed for the determination of ivermectin residues in crucian blood,muscle,hepatopancreas,kidney and gonad tissues.Homogenized crucian tissues were extracted with ethylacetate prior to centrifugation.The supernatants were evaporated to dryness at 45 ℃ with nitrogen.The residues were dissolved with the mixture of methanolwater(4∶1,V/V)and defatted with hexane.After centrifugation,the underlayers were filtrated with 0.22 μm filter and then 20 μL filtrates were detected by reverse phase high performance liquid chromatography with 245 nm detection wavelength.Its method detection limit was 7.5 μg/L and 7.0 μg/kg in plasma and muscle respectively.The good linear range was 0.025-1.00 mg/kg in crucian muscle,hepatopancreas,kidney,gonad tissues and 0.025-1.00 mg/L in plasma.In different tissues,the average withinbatch and the betweenbatch standard deviations of this method are(2.55±1.78)%-(3.99±1.55)% and(4.18±3.97)%-(6.01±2.54)% respectively,with an average recovery of(90.09±6.54)%-(97.77±5.91)%.After a single oral administration of 0.4 mg/kg body weight,we detected ivermectin residues in crucian muscle tissues with this method.When the water temperature is 25 ℃,we found that the freedrug period of crucian muscle tissues should not be less than 15 days.
林茂 , YANG Xianle , JI Rongxing
2010, 34(6):685-690. DOI: 10.3724/SP.J.1231.2010.06836
Abstract:Cytochrome P450 1A(CYP1A)cDNA is used as a probe for evaluation of environmental contaminant.Grass carp(Ctenopharyngodon idellus)liver cell(GCL),kidney cell(CIK),blastula cell(GCB),snout fibroblast cell(PSF)and ovary cell(CO)were exposed to β-naphthoflavone(BNF)to induce CYP1A.The potency of induction was assessed by the ratio of CYP1A cDNA to β-actin(ACT)cDNA derived from semiquantitive PCR.PCR reaction parameters were optimized so that annealing temperature was chosen to be 57 ℃ and cycle to be 30 times.Under such conditions,the CYP1A expression level could be quantitated by Gauss trace ratio of CYP1A/ACT. PCR amplification product of ACT and CYP1A cDNA showed that CYP1A expression level was very low in control grass carp cells while increased in BNFtreated cells significantly.The expression level of CYP1A in BNFtreated GCL,CIK and CO cells had significant difference(P<0.05),which was sorted as GCL>CIK>CO.But that of GCB or PSF cell could not yet be detected.The state of CYP1A expression in grass carp cells was similar to homologous tissues,and the level in homologous tissues of BNFtreated grass carps was sorted as liver>kidney>ovary.The results indicated CYP1A expression level in vitro had certain correlation with that in vivo.This study illustrates the high potential of fish cell lines in ecotoxicology.
Li Jiale , WANG Gui-lin , BAI Zhi-yi , LI Xi-lei
2010, 34(6):691-700. DOI: 10.3724/SP.J.1231.2010.06776
Abstract:A 1 438 bp full length cDNA sequence of β-actin gene from Hypriopsis cumingii was obtained with RT-PCR and rapid amplification of cDNA ends(RACE)technique.It consists of a 92 bp 5′ untranslated region(UTR), a 1 134 bp open reading frame(ORF)and a 257 bp 3′UTR.The translated protein is composed of 377 amino acids,with 41.9 ku molecular weight,and its calculated isoelectric point was 5.3.The amino acid sequence of β-actin in H.cumingii has six specific amino acid residues:Met178,Ser305,Ser321,Pro325,Val331 and Pro346,respectively.In addition,three other particular base sites of amino acid residues were found in these sequences.Similarly,two characteristic amino acid residues of some mollusks were obtained.The amino acids sequence of β-actin in H.cumingiishared the high similarity with Molluscs,Arthropod and Vertebrate animals(98%-99%).Neighbor-Joining(NJ)tree suggested that H.cumingii clustered with Mollusca firstly,and then clustered with Arthropoda,finally clustered with Fish,Amphibians,Mammals. Semi-quantitative RT-PCR analyses showed that the expression level of β-actin gene in H.cumingii was stable in eight tissues:mantle,blood,liver,kidney,stomach,gut,gill and abdominal foot.
MING Jun-chao , XIE Jun , GE Xian-ping , XU Pang , HE Yi-jin , ZHOU Qun-lan , PAN Liang-kun , ZHOU Chuan-peng
2010, 34(6):792-800. DOI: 10.3721/SP.J.1231.2010.06795
Abstract:Macrobrachium rosenbergii,also known as the giant river prawn,the giant freshwater prawn or the Malaysian prawn,is the most important cultivated species of freshwater shrimp in the world.This species(as well as other Macrobrachium)is commercially important for its value as a food source.In China,the production of M.rosenbergii has been increasing in recent year.However,cultured M.rosenbergii in China as well as in other parts of the world have suffered from various stress factors such as ambient temperature,stocking density,physicochemical parameters,transport,and storage. Stress means the sum of nonspecific responses of organisms to various acute stimuli inside and outside.It is a physiological response of any organisms in the long evolution process to extend its adaptation,but excess response,which inhibits immunity,improves the free radical contents,destroys the homeostasis between producing and cleaningup of free radical,and attacks the double bond of polyunsaturated fatty acid in plasma membrane,the last leading to increase the lipid peroxidation contents and lipid peroxidation injury.So these are many disadvantageous impacts on organisms and it causes fish to infect diseases even to die.Therefore,how to alleviate stress response is the research focus in this field.The present research falls on substances,which can strengthen immunity of organisms and alleviate stress,e.g.vitamin C,vitamin E,lipid acid,metal ion,fructose,astacin. Chinese herbal medicines have many merits:natural,highly effective,low toxicity,low side effects,and rich in resources.Chinese herbal medicines have many effective components,such as organic acids,alkaloid,saccharide,volatile oil,wax,glycoside,tannic acid substance and some unknown factors that can elevate fish immunity and enhance the growth.However,few reports have been found in studying the effects of Chinese herbal medicines on the hepatopancreas antioxidative capacity and the level of HSP70 mRNA under high temperature of M.rosenbergii. In view of this, M.rosenbergii were divided into 5 groups randomly.The control were fed with basal diet,the others the treated groups were fed with basal diet supplemented with 0.05%,0.10%,0.20%,0.40% anthraquinone extracts for 10 weeks,respectively.After that,shrimp were stressed by high temperature at 35 ℃ for 48 h continuously.The changes of total antioxidative capacity(T-AOC),catalase(CAT),superoxide dismutase(SOD),nitrogen oxide(NO),malondialdehyde(MDA)and the mRNA level of heat shock protein 70(HSP70)in hepatopancreas before and after the stress were investigated.The results showed that compared with the control,0.05% dose of anthraquinone extract could elevate CAT,SOD activities and reduce MDA concentration,0.20% dose group improved TAOC and NO content and reduced MDA concentration,and 0.10%,0.20% doses groups improved also the relative content of HSP70 mRNA before the stress.After 35 ℃ high temperature stress,T-AOC,CAT activities,SOD activities,and NO concentrations in hepatopancreas were decreasing in all groups of which the control relatively lower and 0.10%-0.20% doses groups were relatively higher,while MDA contents were increasing trend in all groups of which the control was relatively higher and 0.10%-0.20% doses groups were relatively lower.The relative content of HSP70 in hepatopancreas 6 h after the stress in 0.10% and 0.20% doses groups were higher than those of the control.It is concluded that ingestion of 0.10%-0.20% doses of anthraquinones extracts could improve the antioxidative capacity,elevate the relative content of HSP70 mRNA and help prevent negative effects of high temperature stress to a certain degree.