• Volume 37,Issue 4,2013 Table of Contents
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    • Trace elements in the statoliths of jumbo flying squid off the Costa Rica waters

      2013, 37(4):502-511. DOI: DOI:10.3724/SP.J.1231.2013.38077

      Abstract (2608) HTML (0) PDF 5.33 M (2168) Comment (0) Favorites

      Abstract:Jumbo flying squid,Dosidicus gigas,is the most important commercial fishery in east Pacific Ocean,and the statolith in statocyst is widely applied in the field of population identification,life history analysis and habitat environment.In this paper,we studied the trace element composition of statolith in Dosidicus gigas and its relationship with the corresponding sea surface temperature(SST)by laser ablation inductively coupled plasma mass spectrometry(LA-ICP-MS),then deduced the habitat in different life stages.The results showed that 54 kinds of elements were found in statolith of Dosidicus gigas,calcium(Ca),sodium(Na)and strontium(Sr)are the major elements.Sr/Ca ratio in total statolith is higher than that in Todarodes pacificus,Loligo gahiand Gonatus fabricii.Sr/Ca ratio is the highest in embryonic phase and declined by the age,and its relationship with SST shows a reciprocal ratio.There is no significant difference of Sr/Ca ratio between different hatching months in every life stage.Ba/Ca ratio shows a “U”form from larva to adult,this could be seen as the indicator of vertical movement with larva living in the surface and deep layer for adult.Nucleus is higher than peripheral zone in Mg/Ca ratio which presents positive correlation with SST,and this might be possible with the gradua decrease of growth rate in statolith.Meanwhile,this study also shows that Dosidicus gigasin embryonic phase used its own vitellicle as the nutrient substance,so the trace elements in this phase are related with genetic factor in parent rather than the water environment.

    • Establishment of a TaqMan real-time PCR assay for detecting the Cyprinid herpesvirus Ⅱ

      2013, 37(4):607-613. DOI: DOI:10.3724/SP.J.1231.2013.38441

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      Abstract:A 1 446 bp coding region of Cyprinid herpesvirus Ⅱ(CyHV-2)DNA helicase gene was amplified by PCR and cloned into pMD19T vector for the construction of recombinant plasmid.After being identified and confirmed with PCR reaction,10-fold serial dilutions of recombinant plasmid were used as standard templates for TaqMan real-time PCR to quantify the virus genomic copy number and generate standard curve.Herein,a TaqMan real-time PCR of detecting CyHV-2 was developed.It had a good linear relationship between the initial templates and C-t values with a detection range from 1×101 copies/μL to 1×107 copies/μL,the correlation coefficient(R2)was 0.999 1,and the slope value of standard curve was -3.412.The detection results showed that the specificity of this assay was high for CyHV-2 without cross reactions with DNA templates from KHV and GSIV.The diseased crucian carp from Sheyang and Baoying,Jiangsu Province,were detected with the established method and the results showed that the content of CyHV-2 were 6.89×104 copies/μL and 3.02×102 copies/μL,respectively.The real-time PCR assay described here with high sensitivity and accuracy is considered to be a powerful tool for the rapid detection and quantification of CyHV-2 in fish.

    • Primary study on neuronal development of the embryo and early larvae of the mussel Mytilus coruscus

      2013, 37(4):512-519. DOI: DOI:10.3724/SP.J.1231.2013.38433

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      Abstract:In the present study,the development of FMRFamide-,serotonin-containing cells in the mussel Mytilus coruscus was examined using immunocytochemical and histofluorescent techniques.During the embryonic development,no FMRFamide-like immunoreactive(lir)and serotonin-lirsensory cell appeared.Neurogenesis started during the trochophore stage at the apical extreme with the appearance of one FMRFamide-lircell and one serotonin-lir sensory cell.The numbers of FMRFamide-lir cell increased at 36 hours postfertilization(hpf)and appeared on the two sides of apical extreme.Fibres of two lateral FMRFamide-lir apical cells projected posteriorly and reached developing foot.The location of the first FMRFamide-lir apical cells in the most ventral apical region did not change during early development.At 48 hpf,three FMRFamide-lir apical cells,two lateral FMRFamide-lir apical cells and two cells of developing foot appeared.The FMRFamide-lirfibres projected and the basal neuropil formed.The earliest serotonin-lir cell appeared at 18 hpf at the anterior extreme of the apical region of developing apical organ.At 30 hpf and 36 hpf,the serotonin-lir cell extended a short basal fibre into the region of the developing neuropil of the apical organ.The numbers of the serotonin-lir cell increased to 2-3 at 42 hpf and appeared in the apical region of the D-shaped larvae.Four serotonin-lir cells were detected at 48 hpf and these cells projected basal fibres into the compact apical neuropil.

    • Effects of pH and chromatographic column on isolation,purification of antibacterial peptide from Japanese eel liver,and determination of antibacterial activity

      2013, 37(4):614-621. DOI: DOI:10.3724/SP.J.1231.2013.38284

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      Abstract:In order to quickly isolate and purify antimicrobial peptide,different flow phase pH values(cation-change pH 3.0,4.0,5.0,and reverse-phase liquid chromatography pH 2.0,4.5,7.0),and different chromatographic columns(cation-change and gel chromatography coupled with reverse-phase liquid chromatography respectively)were used to isolate and purify antibacterial peptide fromAnguilla japonica liver,and the results were analysed and compared.The merits and drawbacks of agar plate diffusion and microporous liquid culture methods on determination of antibacterial activity were analysed.The results showed that pH 4.0 buffer solution was the best flow phase in three-pH-value buffer solutions,which had 16.43% extraction ratio and isolated two elution peaks;We did not obtain perfect result from reverse-phase liquid chromatography(RPC)in three-pH-value buffer solutions;Sephadex G-25 gel coupled with RPC significantly improved the isolation effect,elution peak amounts were greater,peaks were sharper and narrower,baseline was flatter and low than those of cation exchange coupled with RPC.Agar plate diffuse method to determine antibacterial activity was simple to operate,and experimental result was intuitional.It required much protein,so it should be used to determine antibacterial activity during protein crude extraction period;Microporous liquid culture method to determine antibacterial activity was sensitive.It required little protein,so it should be used to determine antibacterial activity during isolation and purification of protein in chromatography period.

    • Ontogeny of the gill and Na+,K+-ATPase activity of rock bream(Oplegnathus fasciatus)

      2013, 37(4):520-525. DOI: DOI:10.3724/SP.J.1231.2013.38395

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      Abstract:Based on the histological observations under optical microscope,we studied the ontogeny of the gill of rock bream Oplegnathus fasciatus at (24?1.0)℃.At hatching,gill anlage was visible in the pharyngeal region.On 2 DAH(Day After Hatching),as the branchial cavity was formed,four pairs of gill arches were evident and some muscular fibres were observed in gill arches.The blood cells were visible in the vascular structures of gills on 3 DAH,and the primordial filaments first appeared in the second and third arches,then in the first and fourth arches on 4 DAH.The primordial lamellae were first formed in the filaments of the second and third gill arches on 6 DAH,then in the first and fourth arches on 7 DAH.On 6 DAH,the pseudobranch anlage was observed as a paired structure lied by the anterior branchial cavity and the lamellae in the pseudobranch were visible on 7 DAH.The chloride cells were first arranged in the base of the lamellae on 7 DAH,then also in the base of filaments on 8 DAH.The pavement cells were visible in the branchial epithelium on 8 DAH and the pillar cells were developed to delimit the vascular structures of lamellae on 9 DAH.On 10 DAH,the mucous cells were seen in the lamellae of pseudobranch.From 14 DAH forward,the filaments and lamellae of gills increased in number and length prominently and the gill of larvae was similar to that of juvenile fish.The specific activity of Na+,K+-ATPase increased from cleavage period(0.712?0.400)U/g to segmentation period(2.315?0.515)U/g,and then decreased progressively to a low level until 9 DAH(1.389?0.734)U/g.Subsequently,the activity increased.After 12 DAH,it decreased to the minimum at (0.246?0.126)U/g on 14 DAH.Then the activity increased rapidly and reached the maximum at(4.731?0.309)U/g on 18 DAH.From 20 DAH,the specific activity of Na+,K+-ATPase reached a relatively stable level until 50 DAH(3.667?0.633)U/g.The change of Na+,K+-ATPase activity is closely related with the development of gill,especially the chloride cells.The mortality of rock bream larvae was very high in the period when the specific activity of Na+,K+-ATPase reached the minimum from 12 to 16 DAH.

    • Optimization of enzymatic technology of yellow-margined box turtle muscle and antioxidant abilities of its hydrolysates in vitro

      2013, 37(4):622-630. DOI: DOI:10.3724/SP.J.1231.2013.38351

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      Abstract:Antioxidants are known to be beneficial to human health as they may protect the body against damage by reactive oxygen species,which can attack DNA,protein and membrane lipids and play a critical role in a variety of disease such as cancer,diabetes,cardiovascular disease and Alzheimer’s disease.However,there is much evidence that the use of natural antioxidants in foodstuffs is more attractive because of the potential health hazards associated with synthetic antioxidants in vivo.Turtle not only has a high nutritional value,but also has a potential medicinal value.In order to better know turtle’s nutrition and healthy function,the functional properties of enzymatic hydrolysates from yellow-margined box turtle(Cistoclemmys flavomarginata)muscle were investigated in this research.An enzymatic treatment procedure was devised to make the preparation of antioxidant peptide.By using a L9(34)orthogonal array design,the optimum conditions for hydrolyzing yellow-margined box turtle muscle with Alcalase to obtain antioxidant peptides with the highest hydroxyl radical scavenging activity were as follows:pH 8.0,temperature 55 ℃,material to liquid ratio 1.5∶20 g/mL,enzyme-to-substrate ratio 3.0% and reaction time 3 h.Under the above conditions,the scavenging rate of hydroxyl radical was 82.08%.Subsequently,the antioxidant properties of yellow-margined box turtle hydrolysates were determined in vitro.The results showed that the enzymatic hydrolysates had good scavenging abilities on hydroxyl radical(·OH),superoxide anion radical(O2-·),α,α-diphenyl-β-picrylhydrazyl radical(DPPH·)and hydrogen peroxide(H2O2),and they also had certain reducing power,ferrous ion chelating capacity and the inhibition of linoleic acid autoxidation.In general,the yellow margined box turtle hydrolysates showed certain antioxidant activities in different systems in vitro,and they will have new and broad prospects.

    • The changes in lipid and fatty acid profiles of devil stinger Inimicus japonicas during the development of embryo and yolk-sac larvae

      2013, 37(4):526-535. DOI: DOI:10.3724/SP.J.1231.2013.38335

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      Abstract:wild-caught broodstocks of devil stinger Inimicus japonicas were induced with LHRH-A3 to spawn and the fertilized eggs were incubated artificially in seawater with salinity 29 at 20 ℃.The lipid compositions and fatty acid profiles of the embryos(blastula stage to tail bud stage)and yolk-sac larvae[newly hatched,1 day post hatching(DPH),2 DPH,3 DPH(unfed)]were investigated.The results indicated that the total lipid content of devil stinger decreased from 13.85% of the blastula stage embryos to 11.66% of 3 DPH larvae.Polar lipid accounted for 72.20%-75.39% of total lipid during the early development.There was no significant difference in total lipid and polar lipid contents during embryogenesis.The total lipid and polar lipid contents declined significantly during the development of yolk-sac larvae.The neutral lipid content first increased,then decreased significantly during embryogenesis and kept stable during the development of yolk-sac larvae.DHA(22∶6n-3),16∶0,ARA(20∶4n-6),EPA(20∶5n-3),18∶0 and 18∶1n-9 were the dominant fatty acids in total lipid of embryos as well as of yolk-sac larvae.The contents(mg/gDW)of DHA,ARA and EPA in total lipid as well as in polar lipid declined significantly during the early development of the devil stinger.There was sharp declining in contents of DHA and ARA in total lipid as well as in polar lipid when the embryo developed from blastula stage to tail bud stage.The EPA and DHA contents in neutral lipid first increased,then decreased during the early development and the peak appeared at newly hatched larvae stage and 2 DPH larvae respectively.The ARA content in neutral lipid increased step by step during the early development.DHA and ARA to EPA,saturated fatty acids(SFAs)and poly-unsaturated fatty acids(PUFAs)to mono-unsaturated fatty acids(MUFAs)as well as N-6 PUFA to N-3PUFA were preferentially utilized in total lipid during the early embryonic development.While EPA to DHA and EPA to ARA in total lipid were preferentially utilized during the later embryogenesis and development of yolk-sac larvae.Among the SFAs,16∶0 to 18∶0 was preferentially utilized during the whole early development.Among the MUFAs,16∶1 to 18∶1 was preferentially utilized during the later embryogenesis and development of yolk sac larvae.It was therefore suggested that DHA,EPA and ARA in polar lipid could be transferred into neutral lipid during the early development of devil stinger.And the preferential utilization of some fatty acids depended on the lipid class and the development stage of the embryo and yolk-sac larvae.

    • Isolation and identification of ACE inhibitory peptides from the autolysis product of shrimp head(Litopenaeus vannamei)

      2013, 37(4):631-640. DOI: DOI:10.3724/SP.J.1231.2013.38204

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      Abstract:Shrimp head is susceptible to autolysis under certain conditions,the protein in it is degraded into soluble protein,peptides and amino acids,and some peptides are active peptides which can inhibit the ACE enzyme activity.At present,many ACE inhibitory peptides derived from food protein have been developed.In the present study,two ACE inhibitory peptides(Tyr-Pro and Leu-Pro/Ile-Pro)were highly purified from the shrimp head(Litopenaeus vannamei)autolysate by extra fine membrane and a series of column chromatographies.In the first autolysis solution of shrimp head was consecutively extracted through extra fine membrane with molecular weight cut-offs(MWCO)at 8,5,3 ku,respectively.The active results shown that filtrate through MWCO at 3 000 u had the highest ACE inhibitory activity.The crude filtrate through MWCO at 3 ku was purified by Sephadex G-25 gel chromatography,SP Sephadex C-25 anion exchange chromatography as well as Sephadex G-15 gel chromatography,respectively.After that,the ACE inhibitory activity of purified filtrate almost increased by 8 times(IC50=0.19 mg/mL)that of crude filtrate.The high active collected fraction from Sephadex G-15 gel chromatography was carried out by RP-HPLC(High-Performance Liquid Chromatography)twice for the further purification and two kinds of dipeptide were obtained,and the identification of dipeptide by mass spectra showed that they were Tyr-Pro and Leu-Pro/Ile-Pro,and the molecular weight was 279 u and 229 u,respectively.

    • The initial ovarian development and the expression of some related genes in ricefield eel,Monopterus albus

      2013, 37(4):536-544. DOI: DOI:10.3724/SP.J.1231.2013.38468

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      Abstract:The initial ovarian development of ricefield eel Monopterus albus raised under laboratory conditions was investigated with histological analysis,and the expression of gdf9(growth differentiation factor 9),cyp19a1a (P-450 aromatase),amh(anti-M?llerian hormone)and amhrII(anti-M?llerian hormone type II receptor)was analyzed with Real-time PCR assay.From the age of 4 months to 9 months,the oocytes in the ovary of juvenile ricefield eel remained in the primary growth stage,and the basophilic Balbiani’s vitelline bodies(Bb)increased in number at the beginning,then decreased,and Bb’s shape changed from dot at first to large cluster,then to small cluster.At the age of 10 months,the oocytes in the ovary of the juvenile ricefield eel entered the secondary growth stage,with the appearance of cortical alveoli in the cytoplasm and the differentiation of the somatic cells around the oocyte into granulosa and theca cells,and Bb either disappeared or degenerated to dot,indicating that the ovarian puberty of the juvenile ricefield eel was initiated.At the age of 12 months,the ovary of the ricefield eel was filled with late vitellogenic oocytes containing migratory or disintegrated germinal vesicles,and reached maturity.The expression ofgdf 9 was high and did not vary significantly from the age of 4 to 12 months,suggesting its indispensable roles in ovarian development during this period.The expression levels of amh,amhrII,and cyp19a1a increased in ricefield eels at the age of 4 to 7 months,but decreased at 8 months.However,the expression of these genes increased at the age of 10 months,coinciding with the initiation of puberty.Taken together,the results of present study indicated that juvenile ricefield eels initiated ovarian pubertal development at the age of ten months,and might reach ovarian maturation at the age of one year.The expression of amh,amhrII,and cyp19a1a might play important roles in the initiation of ovarian puberty.

    • The influences of silver carp and bighead carp in bio-manipulation pen on the community structure of cladocera in Lake Gehu

      2013, 37(4):545-555. DOI: DOI:10.3724/SP.J.1231.2013.38366

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      Abstract:As a dominant group of zooplankton in fresh water worldwide,Cladocera plays an important role in the material circulation and energy flow of water ecosystem,and it is not only a consumer for small animals and algae,but also food for zooplanktivorous fish.Many current studies have showed that non-classical bio-manipulation,a way to prevent water bloom and improve water environment by controlling fierce fish and stocking planktonivorous filter feeding fish(such as Hypophthalmichthys molitrixand Aristichys mobilis),can be effective.However,to some extent,phytoplankton and community structure of cladocera may be varied with different stocking patterns,fish density and lake type.Lake Gehu,a total area of 164 km2 crossing Wujin and Yixing,is located in the south of Jiangsu province.In recent years,more attention was paid to the researches about the influence of H.ypophthalmichthys molitrixand Aristichys mobilis on lake ecosystems,of which the past domestic studies were mainly focused on aquatic plants,physical and chemical factors,the control of eutrophication and phytoplankton,however,no reports have been made in the bio-manipulation fish pen of Lake Gehu about the influences of abiotic environmental factors and filter-feeding fish on cladocera,and that is our work to fill in the blanks.Thus,from December 2009 to November 2010,a large bio-manipulation fish pen with the stock of silver carp(H.molitrix)and bighead carp(A.mobilis)inside was built to control the cyanobacterial bloom in Lake Gehu,an investigation and laboratory analysis was carried out to compare the variation of the cladoceran community structure between region A outside of the fish pen and regions B,D,C with different ratios of fish(1∶4,1∶2,1∶1.5),which was combined to discuss the influences of bio-manipulation fish pen on cladocera and the effects on improving eutrophic water.Through the identification,32 species of cladocera,belonging to 7 families,14 genera were collected inside and outside of the fish pen where there were no differences between species,and the annual average density and biomass of region A were both higher than any region inside of the fish pen,having a significant difference(P<0.05)with region B,respectively.The main dominant species were Bosmina longirostris,Ceriodaphnia cornuta and Moina sp..In August,the density of Moina sp.in region A had significant differences from region B and D,while 3 regions inside of fish pen had no apparent differences,et al.the results of which indicated that Moinasp.with a larger volume was easier to be preyed by fish than the other two small species.Moreover,some characteristic parameters of water were investigated to depict the correlation between cladoceran and environmental variables.CCA(Canonical Correspondence Analysis)showed that water temperature,permanganate index(CODMn),nitrate nitrogen(NO3-N)and water transparency(SD)were the main abiotic factors affecting the community structure of cladocera.

    • Pollution analysis and characteristics of pollution biology at Dalianhu water source based on zooplankton and benthos

      2013, 37(4):556-564. DOI: DOI:10.3724/SP.J.1231.2013.38360

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      Abstract:Pollution biology indices on zooplankton and benthos were used to analyze the pollution state of different areas of Dalianhu water source,in order to study the pollution characteristics of Dalianhu water source.The evaluation effects of different biological indices on water pollution were considered with its water quality indices,and principal component analysis was used to analyze its pollution characteristics comprehensively.The results showed,during summer and autumn,the range of Shannon-Wiener index(H1)and Margalef diversity index(D)of zooplankton in different areas of Dalianhu water source was 0.3-1.8 and 1.0-10.4,respectively;and the range of Shannon Wiener index(H2)and Goodnight index of benthos(G)was 0.81-1.26 and 0.30-0.88,respectively,which all showed it were in pollution.Correlation analysis results was in comparison with the water quality indices,showing the Margalef diversity index(D)of zooplankton and Goodnight index(G)of benthos can reflect the water pollution state preferably.Combined with the results through principal component analysis,the rank order of pollution state of different areas of Dalianhu water source was water forest>inner river>ponds>external river>lake area.The results showed the lake area of Dalianhu water source had certain ability to purify and protect water,and the pollution was mainly from interior sedimentation of organic matter and pond aquaculture.The ecological restoration measures for Dalianhu water source should include reshaping the terrain,sediment dredging,communicating water network,controlling pond aquaculture,and so on.

    • Effect of vitamin C on growth,immunity and anti-ammonia-nitrite stress ability in juvenile black carp(Mylopharyngodon piceus)

      2013, 37(4):565-573. DOI: DOI:10.3724/SP.J.1231.2013.38322

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      Abstract:Studies were conducted to investigate the effects of dietary vitamin C(VC)on growth,immunity and anti-ammonia-nitrite stress ability in juvenile black carp,Mylopharyngodon piceus(initial average weight 7.27?0.40 g).Six iso-nitrogenous and iso-energetic diets were formulated to supply at dosages of 0(control),16.3,33.9,69.1,137.8 and 277.5 mg/kg vitamin C diet,respectively.After feeding 8 weeks,according to specific growth rate,three group of 0,69.1 and 277.5 mg/kg of black carp were subjected to an ammonia-nitrite stress(20 mg/L)for 24 h.Results indicated that the effective vitamin C content for maximum growth is about 63.0 mg/kg.The vitamin VC levels in liver,muscle and serum were in positive correlation with VC content in diet,and no significant differences were observed among fish fed the diets with equal to or higher than 137.8 mg/kg of vitamin VC.No significant differences in total superoxide dismutase(T-SOD)and malondialdehyde(MDA)were observed among dietary treatments.But VCcould significantly increase the catalase(CAT)and glutathione peroxidase(GSH-px)activity(P<0.05).With the VC content increased,CAT increased at first and then decreased.Glutathione(GSH)and Na+/K+-ATPase(NKA)increased when VC concentration in diets were increased,cortisol(COR)were decreased at first and then increased.After ammonia-nitrite stress,each group of the VC concentration in liver,T-SOD,CAT,GSH-px activity and GSH content in serum and gill NKA activity decreased,serum VC level,COR and MDA content increased.The T-SOD,CAT and NKA activity,GSH,MDA and COR content significantly changed before and after ammonia nitrite stress.Liver vitamin C concentration in fishes fed VC diets significantly decreased after stress,however,there are no significant changes in CAT,NKA activity and GSH,cortisol content compared with control group.T-SOD activity in 69.1 mg/kg vitamin C diet had no significant change before and after stress,but GSH-px activity and MDA content had significantly changed.In 277.5 mg/kg vitamin C group,GSH-px activity and MDA content had no significant change except T-SOD activity and serum VC level.All the results indicated that the effective vitamin C requirement of juvenile black carp for optimal growth might be 63.0 mg/kg diet.Fish have been stressed when put in ammonia nitrite,and VC concentration in diet could improve immunity and anti ammonia-nitrtie stress ability of black carp.

    • Isolation and identification of Bacillus sp.and evaluation of its effect on WSSV disease resistance in Litopenaeus vannamei

      2013, 37(4):574-583. DOI: DOI:10.3724/SP.J.1231.2013.38373

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      Abstract:In order to select WSSV disease-resistant strains,a marine Bacillussp.was isolated and purified from digestive tract from the healthy of Chinese shrimp(Fenneropenaeus chinensis).The morphology and gram stain indicated that the strain is a gram-positive and rod-shaped bacterium,with a single polar flagellum and oval spores.The colony is circular and slightly raised.Identification analyses by the Biolog Carbon Source Utilization,ATB Microbial Identification System,and the fatty acid gas chromatography indicated that the most similar strain in physiological and biochemical characteristics is Bacillus firmus.Phylogenetic analysis with 16S rRNA sequence showed that it has 100% homology with the previously reported Bacillus firmus.The cultured strain PC024 was added to feed by conglutinating to the surface of the pellets and fed to Litopenaeus vannamei.After feeding for 20 d,the shrimp was challenged with WSSV by intramuscular injection to observe the cumulative mortality in 14 d post-challenge.The results showed that the experimental group fed with the strain PC024 had a relative survival rate of 33.7% in comparison with the control group.The immune-related enzyme activity in the serum and hepatopancreas of shrimp in the experimental group was significantly increased than the control group.And the total number of bacteria of the intestine of the experimental group is always significantly higher than that of the control group and Bacillus firmus can be isolated from the experimental group.This study suggests that the Bacillus firmusPC024 can be used as the WSSV disease prevention probiotic strains and can further be used in shrimp farming.

    • Selection of the families with high growth rate and high female proportion in half-smooth tongue sole(Cynoglossus semilaevis)

      2013, 37(4):481-488. DOI: DOI:10.3724/SP.J.1231.2013.38328

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      Abstract:The present study aims to uncover the relationship between the phenotypic sex and genotypic sex and the growth rate of male and female half-smooth tongue sole and to select the excellent families with high female proportion.In this paper,we firstly established 22 half smooth tongue sole families which were cultured in the same pond.Secondly,we identified the offspring’s phenotypic sex and genotypic sex of 5 normal male families and 5 neo-male families.This study shows that there are significant differences not only in growth rate but also in proportion of male to female.Two fast growing families(No.16 and No.61)whose relative weight gain rate were 0.81 g/d and 0.56 g/d and 4 relatively fast growing families(No.21,No.28,No.57 and No.63),the relative weight gain rate ranges from 0.48 g/d to 0.53 g/d were selected;12 commonly growing and 4 slow growing families were selected also.The average ratios of genotypic female and phenotypic female of the 5 normal male(ZZ)families were 51.87% and 42.94%,and the ratio of genotypic female reversing to phenotypic male was 14.08%.Instead,the average genotypic female and phenotypic female ratio of the 5 neo male(ZW)families was 49.34% and 4.45%,respectively,and the ratio of genotypic female reversing to phenotypic male was 91.41%.In the present study we found for the first time that there were significant differences in phenotypic sex ratio between different families.We also found that the genotypic females in neo male families were more easily reversed to phenotypic males,which increased the number of phenotypic males in cultured populations.It initially showed that the families with high female proportion and fast growing rate could be selected via family selection,which could provide theories and new technical methods for the breeding and the production of high quality seeding of half smooth tongue sole.

    • Cloning and expression analysis of transcriptional regulator rovS gene of Stretococcus agalactiae ZQ0910 isolated from tilapia

      2013, 37(4):584-592. DOI: DOI:10.3724/SP.J.1231.2013.38348

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      Abstract:Cloning and expression analysis of transcriptional regulatorrovS gene of Stretococcus agalactiae ZQ0910 isolated from tilpia were studied in this study.A pair of primers were designed based on rovS gene sequence published in GenBank.The gene sequence of coding the RovS protein was cloned and then inserted into the pET-28(+)vector to construct prokaryotic expression plasmid pET-28-RovS.The recombinant RovS fusion protein was overexpressed in E.coli Rosetta(DE3)cells in the presence of isopropyl-β-thiogalactopyranoside(IPTG).Sequence analysis revealed that rovS gene is 849 bp and encodes a putative protein of 282 amino acids,and the amino acid sequence of RovS of S.agalactiae ZQ0910 showed highest identity of S.agalactiae 2 603 V and ATCC13813.The His-RovS fusion protein with 34 ku molecular mass was successfully expressed inE.coli Rosetta(DE3).The soluble recombinant protein was highly expressed under induction conditions of exposure to IPTG and successfully purified on Ni2+-IDA column.The purified fusion protein was injected into New Zealand white rabbits to produce polyclonal anti-RovS serum.The antibody titer detected by ELISA reached 1∶512 000.In summary,we identified and characterized a novel regulatory gene,termed rovS,encoding a regulator of virulence in S.agalactiae,which is involved in the expression of known and putative virulence genes in the bacteria.

    • Combining ability of hybrid generation from the introduced populations of Litopenaeus vannamei

      2013, 37(4):489-495. DOI: DOI:10.3724/SP.J.1231.2013.38339

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      Abstract:Seven populations of Litopenaeus vannamei were collected to evaluate the general combining ability and the specific combining ability in harvest body weight and survival with the mixed linear model and generalized linear models,combined with constrained maximum likelihood method.Incomplete diallel crosses and nest design were adopted and 130 full-sib families were obtained by artificial insemination.The results showed that the populations of UA5,UA4 and SIN were higher than others in general combining ability of harvest body weight,the magnitude were 0.40,0.32 and 0.19,respectively.The general combining ability of harvest body weight and survival in the hybrid groups of UA5×SIN,UA1×UA2,UA4×UA5,UA4×SIN and UA1×SIN,which were priority combinations in producing superior offspring,had positive advantages.The analysis of variance of combining ability showed that in different environments,the proportion of GCA and SCA variance component in body weight traits had little difference.Correlation analysis showed that there is positive correlation between the GCA of harvest weight,but negative correlation in survival.For the study of general combining ability and specific combining ability in different groups,the experiment provided the basic data for further genetic breeding of L.vannamei.

    • Recombinant expression and antimicrobial activity analysis of macrophage inflammatory protein-3 α in Mauremys mutica

      2013, 37(4):593-598. DOI: DOI:10.3724/SP.J.1231.2013.38304

      Abstract (2319) HTML (0) PDF 5.34 M (1745) Comment (0) Favorites

      Abstract:Macrophage inflammatory protein-3α is a kind of inducible secreted protein which plays an important role in the immune response and inflammation damage in many diseases through the corresponding receptors on a variety of immune cells to produce chemokines.We obtained the ORF of MaCCL20 by RT-PCR,inserted into the expression vector pET-32a,and then transformed into E.coli BL21.We expressed the fusion protein pET-32a-CCL20 after IPTG induction,prepared the polyclonal antibodies and did research on Western blot and antimicrobial activity of it.The experiments show that the fusion protein is highly expressed in the conditions of 37 ℃,0.8 mmol/L IPTG for 4 hours.The SDS-PAGE gel electrophoresis figure shows that the relative molecular weight of the fusion protein is 30 ku which is the same as predicted before.We got a single band after His Bind nickel column purification indicating the high purity of MaCCL20 recombinant protein.The antibacterial experiments show that MaCCL20 recombinant protein has strong antibacterial effects on Staphylococcus aureus,Serratia marcescens,and Aeromonas hydrophila.

    • Sagittal otolith microstructure,daily age and early growth of Coilia mystus juveniles in the Yangtze Estuary

      2013, 37(4):496-501. DOI: DOI:10.3724/SP.J.1231.2013.38361

      Abstract (2683) HTML (0) PDF 5.74 M (1957) Comment (0) Favorites

      Abstract:Coilia mystus is a commercially important fish in the Yangtze Estuary.Annual capture averaged 2 000 tonnes in the 1980s,accounting for almost half of the total catch in this estuarine area.However,this species has declined dramatically since the late 1990s.C.mystusjuveniles were sampled from the Yangtze Estuary using trawls and set nets in July,August and September 2007.Sagittal otoliths were analyzed to estimate the daily age and early growth of the juveniles.Standard length(LS)of C.mystusjuveniles was between 25.6 and 63.2 mm.Otolith shape of C.mystusjuveniles was similar to that of adults:anomalistic and elliptic,with a single rostral protrusion.There was one nucleus and one primordium on the transverse plane of the otolith.Primordium diameters were between 12.5 and 22.8 μm,averaging (14.3?4.8)μm(n=111).The distance from the central nucleus to the first increment was (24.85?6.38)μm(n=44).The increments in the long axis were thin and unclear,but those in the short axis were thick and clear.Estimated ages of collected specimens ranged from 54 to 128 days.Hatch dates fell between 10 April and 23 July 2007,with the peak between 11 may and 26 June.The relationship of otolith radius(R)to standard length(LS)was fitted to a linear function:LS=6.68+0.065R(r2=0.81,n=111,P<0.05).The incremental width was narrow and decreasing in the first 20 increments;after that,the incremental width increased to the 50th increment,and then remained constant.

    • Studies on the hemocytes types and their immunological functions in bloody clam(Scapharca broughtonii)

      2013, 37(4):599-606. DOI: DOI:10.3724/SP.J.1231.2013.38321

      Abstract (2730) HTML (0) PDF 48.13 M (1700) Comment (0) Favorites

      Abstract:It was the first time to study the types and immunological functions of blood cells in bloody clam(Scapharca broughtonii)by the means of microscopic observation and immunostimulation with pathogenic Vibrio anguillarum,and three types of hemocytes were recognized:red blood cells,white blood cells and thrombus cells.Red blood cells had four developmental states:freshmen,mature,aging and death.White blood cells were divided into acidophils,neutuophils,basophils,lymphocytes,and megalokaryocytic.Acidophils,neutuophils,basophils and thrombus cells were all involved in the blood coagulation.Both red blood cells and white blood cells had phagocytic function,red cells showed the passive phagocytic function and immune adherence function,while white cells showed the active phagocytic function.Acidophils and neutuophils cells were principal in phagocytizing pathogens,and basophils cells could be found phagocytizing large size foreign materials,an increase in the number of white cell and inflammation took place after the bloody clam was infected by pathogen.It was also the first time to show the thrombus cells and its coagulation function in bloody clam(Scapharca broughtonii).The results showed that the blood cells had systemic immunologic defense mechanisms against the mass pathogens invasion into the body of bloody clam(Scapharca broughtonii).

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