• Volume 38,Issue 12,2014 Table of Contents
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    • Effects of inbreeding on growth and survival of larval and juvenile Pacific oyster(Crassostrea gigas)

      2014, 38(12):2005-2011. DOI: 10.3724/SP.J.1231.2014.49358

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      Abstract:The effects of inbreeding on growth and survival of Pacific oyster(Crassostrea gigas)were investigated using inbreeding families which were established by first generation of full-sib mating(F1,F=0.250)and second successive generation of full-sib mating(F2,F=0.375).Control group(F=0)was produced by random mating of wild oysters.The fertilized rate,hatching rate,survival rate,growth and inbreeding depression coefficients were compared between different experimental groups.The results indicated that all the three experimental groups had high fertilized rates ranging from 94.42% to 98.46%,and hatching rates were all above 90% except group F2.The growth of F1 and F2 showed significant inbreeding depression after 12 days of breeding at larval stage.An increasing inbreeding depression of growth was observed with the increase of inbreeding coefficient in group F1 and F2 at post-larval stage.The inbreeding depression coefficient of average shell height was found to be -0.386 0% at F=0.250,-0.647 1% at F=0.375 at 20 d.The inbreeding depression of larval survival was obvious and increasing with age.The inbreeding depression coefficients of larval survival in group F1 and F2 ranged from -0.197 2% to -2.490 5% and -0.136 3% to -2.232 5%,respectively.A significant inbreeding depression in average shell height of spat was observed all the time in families with F=0.250 and F=0.375.Inbreeding depression of shell height of group F2 was higher than group F1 at juvenile stage.However,no significant difference of average shell length of spat was found between control group and the other two inbreeding groups.Inbreeding depression of survival existed all the time in inbreeding families with F=0.250 and F=0.375 at juvenile stage,and both of them increasing with age.This study demonstrates that inbreeding has negative effects on the economic traits,especially on growth and survival.These effects even still exist in the family with second successive generation of full-sib mating,thus it is necessary to highlight the importance to maximize the genetic diversity in selective breeding programs.These results also emphasize the significance of maintaining pedigree records which are used to avoid the deleterious effects of inbreeding depression in shellfish breeding programs.

    • Effects of cholinoceptor compounds on larval metamorphosis of the mussel Mytilus coruscus

      2014, 38(12):2012-2017. DOI: 10.3724/SP.J.1231.2014.49448

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      Abstract:The metamorphosis responses of mussel(Mytilus coruscus)larvae to cholinoceptor compounds were investigated through a series of bioassays in order to obtain the information on muscarinic and nicotinic cholinoceptor in mediating larval metamorphosis in this species.In the inducing effect of the agonist,pediveliger larvae were exposed to the test solution either in 24-h exposure bath or in continuous exposure throughout the experiment.In the inhibitive effect of the antagonist,pediveliger larvae were exposed to the test solution in the absence or presence of the agonist.In the 24 h exposure assays,carbamylmethylcholine induced larval metamorphosis at 10-5 to 10-4 mol/L concentrations,while acetylcholine showed no inducing activity.In continuous exposure assays,carbamylmethylcholine showed inducing activity at 10-5 to 10-4 mol/L concentrations,and acetylcholine induced larval metamorphosis at 10-6 to 10-4 mol/L concentrations.Nicotinic cholinoceptor antagonist hexamethonium bromide did not inhibit larval metamorphosis in the presence of carbamylmethylcholine or acetylcholine.In contrast,muscarinic cholinoceptor antagonist atropine,inhibited larval metamorphosis in the presence of carbamylmethylcholine.In addition,no larval mortality was observed at all test concentrations.Thus,carbamylmethylcholine and acetylcholine can be used as non-toxic and useful inducers of larval metamorphosis in this species,and can improve M.coruscus larval production for aquaculture.The present study provides a novel insight into the mechanism modulating the metamorphosis of larval of the mussel M.coruscus.

    • Effects of light intensity on spore germination,early development of the sporelings of Scytosiphon lomentaria and the dynamic change of epiphytic algae

      2014, 38(12):2018-2028. DOI: 10.3724/SP.J.1231.2014.49457

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      Abstract:In this research,by using experimental ecology to imitate the culture conditions of sporelings of Scytosiphon lomentaria,the filaments of S.lomentaria and the natural sand filtered seawater were used as the experimental material and the culture solution,respectively.Different light intensity which includes 7.2,18.0,27.0,36.0,45.0,54.0,72.0,127.0 μmol/(m2·s)were set to study the effect of light intensity 7.2-126.0 μmol/(m2·s)on spore germination,early development of sporelings of S.lomentaria and the dynamic change of epiphytic algae.The results indicated that:(1)27.0-72.0 μmol/(m2·s)was the appropriate light intensity range for the spore germination of S.lomentaria,and the spore germination rate reached the maximum under the condition of 45.0 μmol/(m2·s),the germination rate was 44.44% after 16 days of the spores releasing.(2)In the natural sand filtered seawater,36.0-54.0 μmol/(m2·s)was the appropriate light intensity range for the early development of sporelings,and the optimal light intensity was 45.0 μmol/(m2·s),under whose condition the epiphytic algae density was the lowest and the epiphytic algae density was 38.4×104 ind/cm2 after 34 days of the spores releasing.(3)In this study,a total of 29 taxa which belong to 13 genera of 2 phyla were identified,and the main dominant species were Nitzschia frustulum,Pseudo-Nitzschia sicula,Oscillatoria laetevirens,Chroococcus turgidus,Auricula sp. and Nitzschia closterium.Nitzschia frustulum showed exponential growth trend under the condition of 7.2-18 μmol/(m2·s),while Auricula sp. and N.closterium presented exponential growth trend under the condition of 27.0-126.0 μmol/(m2·s).In conclusion,45.0 μmol/(m2·s)was the optimal light intensity for spore germination and early development of sporelings of S.lomentaria.The epiphytic algae,which should be controlled in the process of sporelings production,were Auricula sp. and N.closterium.

    • Effects of compound feed and live feed on the growth,survival,and the activities of digestive enzyme,nonspecific immunity enzyme, metabolic enzyme,and antioxidant enzyme of young fish Coilia nasus

      2014, 38(12):2029-2038. DOI: 10.3724/SP.J.1231.2014.49309

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      Abstract:Through the investigation and analysis of the growth,survival rate,and the activities of digestive enzyme,nonspecific immunity enzyme,metabolic enzyme,and antioxidant enzyme of young fish Coilia nasus,which were fed with compound feed and live feed for 178 days,the effects of compound feed and live feed on growth,survival,and some enzyme activities of young fish C.nasus were studied.The results show that the final body length,final body weight,survival rate,condition factor,and hepato somatic index of compound feed group(125.17 mm,6.27 g,65.73%,0.31 g/cm3,and 1.4%,respectively)were significantly lower than those of live feed group(150.66 mm,12.39 g,85.59%,0.36 g/cm3,and 1.9%,respectively),while there was no significant difference between the two groups in the ratio of intestine length to body length(25.3% and 23.6%,respectively);No protease was detected in livers of two groups,pyloric caecum alkaline protease activity of compound feed group(43.49 U/mg prot)was significantly lower than those of live feed group(86.37 U/mg prot),while there was no significant difference between the two groups in activities of stomach acid protease and intestine alkaline protease;Amylase activities in intestine and pyloric caecum of compound feed group(196.63 and 575.93 U/g prot,respectively)were significantly lower than that of live feed group(928.91 and 1 755.90 U/g prot,respectively),while there was no significant difference between the two groups in amylase activities in liver and stomach;No lipase was detected in livers and stomachs from compound feed group and live feed group,intestine lipase activity of compound feed group(23.55 U/g prot)was significantly higher than that of live feed group(14.39 U/g prot),while there was no significant difference between the two groups in lipase activity of pyloric caecum(17.90 and 13.23 U/g prot,respectively);Alkaline phosphatase(AKP)activity in liver of compound feed group(103.44 U/g prot)was significantly higher than that of live feed group(58.20 U/g prot),while glutamic oxalacetic transaminase(AST/GOT)in liver of compound feed group(20.38 U/g prot)was significantly lower than that of live feed group(32.51 U/g prot).In addition,there was no significant difference between the two groups in activities of acid phosphatase(ACP),glutamic-pyruvic transaminase(ALT/GPT),superoxide dismutase(SOD),glutathione peroxidase(GSH-PX),and catalase(CAT)in liver,and activities of glutamic-pyruvic transaminase(ALT/GPT),glutamic oxalacetic transaminase(AST/GOT),superoxide dismutase(SOD),and glutathione peroxidase(GSH-PX)in serum.The results suggest that C.nasus could ingest the compound feed,there was no significant difference between compound feed group and live feed group in most of digestive enzyme,nonspecific immunity enzyme,metabolic enzyme,and antioxidant enzyme activities,while the growth and survival rates of compound feed group were significantly lower than those of live feed group.Therefore,in order to gradually replace live feed,optimized compound feed formula and improved compound feed that are suitable for C.nasus are recommended.

    • Effects of silymarin on hepatic DNA damage and expression of related cytokines in Cyprinus carpio var.jian

      2014, 38(12):2039-2048. DOI: 10.3724/SP.J.1231.2014.49431

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      Abstract:The present study aimed to evaluate the hepatoprotective effects of silymarin against carbon tetrachloride(CCl4)-induced hepatotoxicity in Cyprinus carpio var.jian.Fish were fed diets containing four doses of silymarin(0,0.1,0.5,and 1.0 g/kg)for 60 d,and then given an intraperitoneal injection of 30% CCl4 in olive oil at a volume of 0.05 mL/10 g body weight.At 72 h post injection,liver samples were collected for the liver index,comet assay,histopathological examination and cytokines mRNA detection.Results showed that the increase of the liver index induced by CCl4 was significantly inhibited by pre-treating the fish with 0.5 and 1.0 g/kg silymarin in the diets.Comet assay showed silymarin could effectively reduce DNA fragment generation of hepatic cells;tail moment,olive tail moment,tail length and tail DNA% were positively changed in fish pretreated with 0.5 and 1.0 g/kg silymarin.In the liver tissue,silymarin had a protective effect on Jian carps with the alleviation of histological changes such as cell swelling,extensive vacuoles degeneration,karyopyknosis and karyolysis.At the same time,the elevation of nuclear factor-κB(NF-κB/C-Rel)and iNOS mRNA expressions induced by CCl4 were also inhibited by the pre-treatments with 0.5 and 1.0 g/kg silymarin,and the IL-1β mRNA expression was significantly inhibited by pre-treatment with 0.1,0.5 and 1.0 g/kg slymarin.The positive effects of silymarin on the liver index,comet assay,histopathological examination and gene expressions were observed in a dose-dependent manner.Overall results prove that silymarin can improve the degree of hepatic cell DNA damage,and the hepatoprotective functions may be related to inhibition of the NF-κB activation and down-regulation of the downstream genes expression such as iNOS and IL-1β.

    • Molecular cloning and expression patterns of the cytochrome CYP17-I gene during the reproductive cycle in Navodon septentrionalis

      2014, 38(12):1945-1955. DOI: 10.3724/SP.J.1231.2014.49392

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      Abstract:As a key steroidogenic enzyme,cytochrome P450c17(CYP17,17α-hydroxylase/17,20-lyase)plays a critical role in the production of sex steroid hormone in many teleost.In order to evaluate the functions of P450c17-I during the gonad development and the relationship between its expression and sex steroid hormone levels,we first cloned and characterized the full-length cDNA of P450c17-I in Navodon septentrionalis.The full-length cDNA of CYP17-I is 1 580 bp,encoding 511 amino acids.The analysis of the structure shows that CYP17-I contains relatively conserved sequence.Then RT-PCR method was used to check the expression of the CYP17-I in different tissues.The result shows that the higher expressions are in ovary,testis and brain,there is relatively low expression in heart,spleen,kidney,and there is much lower expression in pituitary and gill.In addition,the method of 125I labeled radioimmunoassay(RIA)is applied to examine the changes of serum testosterone(T),17β-Estradiol(E2)levels during the reproductive cycle.The results show that the female fish has the highest level of E2 at the early stage of vitellogenesis and it decreases along with spawning.In the male fish the level of E2 was not significantly different in the reproductive cycle.But the level of T in both male and female fish was developmental stage-dependent and the highest level of T appear at 5th month.RT-PCR showed that the expression patterns of CYP17-I were developmental stage-dependent,which is consistent with the changes of T in male fish and female fish.And the highest expressions of CYP17-I are found in the gonad of both genders before the pre-ovulation phase in the reproductive season.But there is no obvious relationship between E2 and CYP17-I expressions.These suggest that CYP17-I plays a critical role in the production of male testosterone hormone in N.septentrionalis.

    • Effects of dietary yeast extract supplementation on the immune-related gene expressions and vibrio-resistant ability in Litopenaeus vannamei

      2014, 38(12):2049-2058. DOI: 10.3724/SP.J.1231.2014.49326

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      Abstract:The basal diet containing 24.5% fish meal as control,and experiment diet(YE group)with 2.5% yeast extract in basal diet,were fed to the white shrimp Litopenaeus vannamei respectively for 56 d,then the mRNA expression levels of Toll receptor,IMD and lysozyme in gills and Vibrio-resistance of the both shrimp were detected for assessing the effect of dietary yeast extract on sensitivity and balance of immune-related gene expression of shrimp.The results showed that:The shrimp fed the experiment diet displayed significantly increased mRNA expressions of Toll receptor and lysozyme in gills compared with the shrimp fed the control diet(P<0.05).There was no significant difference in IMD mRNA expression level in gills between YE group and the control(P>0.05).The relative mRNA expression levels of Toll receptor,IMD and lysozyme of the two groups changed significantly after Vibrio alginolyticus challenge,and the peak of the gene expressions appeared at 24,42 and 36 h respectively in both groups.The peaks of the gene expression of Toll receptor,IMD and lysozyme in YE group were 201.06%,481.46% and 276.77% of those in the control,and significantly higher than those in the control(P<0.05).The mRNA expression levels of Toll receptor and IMD in YE group significantly up-regulated at 12 and 24 h respectively while at 24 and 42 h respectively in the control after Vibrio challenge.There was no significant difference in 72 h cumulative mortality after V.alginolyticus infection between the two groups.It is therefore suggested that yeast extract added in diet significantly up-regulated the relative mRNA expression levels of Toll receptor and lysozyme in gills of L.vannamei before V.alginolyticus challenge,led to significant up-regulations of the relative mRNA expression levels of Toll receptor and IMD early and increased the peaks of the relative mRNA expression levels of Toll receptor,IMD and lysozyme in gills of the shrimp after V.alginolyticus challenge.This result implied that dietary yeast extract improved the sensitivity of the immune related genes expression of the shrimp to some extent.

    • Construction of transgenic P0 grass carp by capsid-targeted viral inactivation of reovirus

      2014, 38(12):1956-1963. DOI: 10.3724/SP.J.1231.2014.49402

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      Abstract:To explore new methods to prevent the grass carp(Ctenopharyngodon idella)hemorrhage,the capsid-targeted viral inactivation(CTVI)strategy was developed in the present study.By combination with Cyprinus carpio heat shock protein 70 or Xenopus EF1α promoters,two different transgenic plasmids pTgf2-EF1α-VP3-SN and pTgf2-Hsp70-VP3-SN were constructed harboring with efficient Tgf2 transposon element.Both transgenic plasmids contain open reading frame(ORF)fusion with GCRV capsid protein VP3 and Staphylococcus aureus nuclease(SN).The transgenic grass carp was produced by co-injection pTgf2-EF1α-VP3-SN or pTgf2-Hsp70P-VP3-SN plasmids into 1-2 cell fertilized eggs with in vitro synthesized Tgf2 transposase mRNA.The PCR and sequencing analysis have proved that the anti-GCRV transgenic systems have successfully been integrated into the genome of grass carp.The transgene positive rates of pTgf2-EF1α-VP3-SN or pTgf2-Hsp70P-VP3-SN plasmids are 40.2% and 37.0%,respectively.Our results demonstrated Tgf2 transposon can efficiently mediate transgenesis in grass carp for fusion ORF of GCRV capsid protein VP3 and SN.Total 120 transgenic grass carp individuals have been obtained in the present study.The construction of anti-GCRV transgenic P0 will provide the materials for future transgenic breeding to prevent grass carp hemorrhage.

    • Studies on mutation breeding of an amylase-producing marine actinomycete strain and its fermentation conditions

      2014, 38(12):2059-2067. DOI: 10.3724/SP.J.1231.2014.49344

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      Abstract:To obtain amylase from marine source,we bred a strain Streptomyces roseofulvus A46 screened from intertidal zone of Zhoushan by ultraviolet and DES mutation.The enzyme activity reached 105.6 U/mL by flask culture,which was 213.3% higher than that of the parent strain,A46.Descent of only 3.6% in enzyme activity of the mutant was observed after ten times of subculture,and the obtained mutant is a relatively stable strain.The optimized fermentation conditions were formulated using Plackett-Burman design and response surface analysis(RSA):culture temperature was 33 ℃;inoculum size was 2.25%;salinity was 23.4;initial pH was 8.0;rotating speed was 200 r/min;medium volumes was 20%;starch content was 0.5%;incubation time was 72 h.Researches show that the enzyme activity was 125.8 U/mL under the optimum condition,which was 19.1% higher than that of non-optimized condition.

    • Effect of KK-42 on the carapace structure in Macrobrachium nipponense during premolt D3 stage

      2014, 38(12):1964-1969. DOI: 10.3724/SP.J.1231.2014.49337

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      Abstract:In order to research the possible mechanism that the duration of molting cycle of the juvenile prawn Macrobrachium nipponense was shortened by KK-42,the cuticle structure of carapace was investigated by the histological method and the N-acetyl-β-D-glucosaminidase(NAGase)activity from hepatopancreas was quantitatively analyzed using ultraviolet absorption method before and after KK-42 treatment.The prawns with body length of (3.3±0.5)cm were randomly divided into two groups:KK-42 treatment group and control one.The prawns were soaked for 1 min in KK-42 solution at a concentration of 1.95×10-4 mol/L(KK-42 treatment group)or 0 mol/L(control group),respectively.Afterwards,the carapaces derived from late premolt(D3)prawns were chosen to be used for histological observation and thickness measurement of layered exoskeleton,meanwhile,the hepatopancreas NAGase activity in stage D3 M.nipponense was quantitatively analyzed.The results showed that the exocuticle thickness of carapace significantly increased by 42.83% compared with the corresponding control on 1st day after KK-42 treatment,and the endocuticle thickness as well as the whole exoskeleton thickness significantly increased by 70.72%(P<0.01),77.27%(P<0.01)and 38.69%(P<0.01),52.23%(P<0.01),respectively,compared to the corresponding control on the 3rd day and the 9th day after KK-42 treatment.A 31.69% rise in the height of epithelial cells was obtained on the 3rd day after KK-42 treatment compared with the corresponding control.The NAGase activity significantly increased by 42.53%(P<0.01),34.28%(P<0.01)和18.36%(P<0.05),respectively,at 6 h,12 h and 24 h after KK-42 administration compared to the corresponding control.The results reveal that KK-42 treatment can significantly increase the cuticle thickness of carapace,and promote NAGase activity from hepatopancreas.

    • Detection of pathogenic vibrios infection in Larimichthys crocea and its forecast and warning of disease

      2014, 38(12):2068-2074. DOI: 10.3724/SP.J.1231.2014.49404

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      Abstract:In order to prevent and control the incidence of ulcer in large yellow croaker,we evaluated the prevalence of infection with three pathogenic vibrios in cultured Larimichthys crocea in Xiangshan by specific multiplex PCR.We collected the liver,kidney,spleen,muscle of fish in each sentinel cage randomly and detected the infection status.The results showed that there were some differences between the three kinds of pathogenic vibrios that infected the fish.The infection rate of V.alginolyticus and V.harvey large in L.crocea was higher than V.parahaemolyticus.The three vibrios were more susceptible to muscle,spleen than to the liver and kidney.The infection rate of different age of fish was not identical.In every sampling time,there existed infection rate.From July to September was the peak of infection rate and the rate increased after the typhoon certainly.In addition,according to the investigation on the relationship between incidence of ulcer and the prevalence of pathogenic vibrio,it showed that the infection rate would reach a higher value a few days before the outbreak of ulcer disease.So we could forecast the incidence and prevalence of the diseases through molecular epidemiology investigation and analyzing the infection of three pathogenic vibrios in the fish.Then,we will establish RT-PCR method of three vibrios to analyse the intensity of infection further.Thus the experimental results will be more convincing.

    • A microsatellite panel for triploid verification in the Pacific oyster(Crassostrea gigas)

      2014, 38(12):1970-1975. DOI: 10.3724/SP.J.1231.2014.49325

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      Abstract:In the Pacific oyster(Crassostrea gigas),triploidy induction is the most common genetic method to enhance production yield through phenotypic improvement.Production of triploids necessarily requires a proper method to verify the ploidy level after induction.Various methods have been developed such as flow cytometry,chromosome counting by karyotype analysis,counting of nucleoli,particle size analysis and so on.However,each method has some drawbacks considering size of sample,accuracy,complexity in conduction and cost.This study aims to identify a microsatellite panel for triploid validation in the Pacific oyster and further explore the number of microsatellites required to successfully verify ploidy status.Triploid oysters are usually induced by suppressing the extrusion of the second polar body.If the distance between a microsatellite and centromere is great,homologous chromosomes might cross over at prophase I of most meiosis allowing the triploid progenies having both of the two maternal alleles.This study screened the 56 microsatellites located in the centromere mapping and obtained seven successfully amplified loci at which the female parent is heterozygous and does not share any allele with the male parent.Using these seven microsatellites,40 triploid oysters were verified from the 115 individuals treated by cytochalasin B(CB,0.5 mg/L)for 15 min starting from the time when about 50% of the eggs released the first polar body.Ploidy status was then verified by flow cytometry and the correspondence was 100%.The ability of a microsatellite in identifying triploids relies on its frequency of microsatellite-centromere recombination(y).To further explore the number of microsatellites required to successfully identify all the triploids,different combinations of microsatellites were analyzed.The results indicate approximate 100% accuracy when product of(1-y)of all microsatellites is no more than 0.005.This study shows that microsatellite markers can serve as an accurate,fast,cost-effective and technically simple method for triploid verification in the Pacific oyster.The protocol described in this work will help in the application of triploids verification using molecular markers in other species.

    • Effect of different hydrolyzed ferrous chelates treatment on iron-deficiency anemia in rats

      2014, 38(12):2075-2083. DOI: 10.3724/SP.J.1231.2014.49397

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      Abstract:This paper focused on the high efficient utilization of by-product of Trichiurus haumela.The hydrolyzed ferrous chelate prepared from by-product of T.haumela was prepared and initially classified by the method of gel chromatography,and recovery and molecular weight(MW)were confirmed.The constitutes and content of amino acid of different chelates were investigated using automatic amino acid analyzer,while the amino acid score were also confirmed.The therapeutic effect of different molecular weight chelates in improving iron-deficiency anemia was conducted by IDA model rats.The results showed that the chelation was classified into 4 materials,the molecular weight of which were 35 400,8 720,465 and less than 200 u,respectively.All of these materials belonged to incomplete protein except component Ⅱ.The evaluation results of amino acid model of ideal protein showed that AAS of these 4 materials were 85.31%,88.22%,101.32%,105.83%,and were also high quality protein.Content of Fe differed significantly from each other,the content of Fe of component Ⅰ and Ⅱ were 5.23% and 3.34%,and there was a similarity in Fe content between Ⅲ and Ⅳ(4.22% and 4.58%).The results of rat experiment showed that different chelation did not have the same effect on weight,serum iron,TIBC,liver MDA and SOD,and component Ⅱ showed the optimal effect in anemia improvement compared with FeSO4.Though component Ⅰ did not possess the significant effect,it also had a certain ability in curing anemia.

    • Gene cloning and expression analysis of MnSOD and CAT from Ulva prolifera

      2014, 38(12):1976-1984. DOI: 10.3724/SP.J.1231.2014.49301

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      Abstract:Manganese superoxide dismutase(MnSOD)and catalase(CAT)are key enzymes in the antioxidant system and important indicators in the resistance study of plants.In this paper,partial cDNA fragments of MnSOD and CAT genes were cloned,and their expression patterns in response to different temperatures and salicylic acid concentrations were investigated by real-time quantitative PCR technique in macroalga Ulva prolifera(Chlorophyta).The cloned MnSOD cDNA was 318 bp,which encoded 105 amino acid residues.The putative MnSOD amino acid sequence of U.prolifera shared 89% identity with that of U.fasciata.Total 229 bp CAT cDNA was obtained,which showed 99% identity with that of U.fasciata,and 93% identity with that of Haematococcus pluvialis by Blast analysis.The phylogenetic trees suggested that U.prolifera clustered with that of U.fasciata in the tree constructed with MnSOD amino acid sequences;and U.prolifera also clustered with U.fasciata firstly,and then clustered with H.pluvialis and Chlamydomonas reintmrdtii in the tree constructed with CAT amino acid sequences.The expression of MnSOD and CAT genes varied in response to different temperatures,of which high temperature showed the most significant effect on the two genes' expression.The expression of MnSOD and CAT was 2.18-fold and 2.05-fold in 35 ℃ culture condition compared with that at 25 ℃,respectively.The expression of MnSOD and CAT was 1.47-fold and 1.31-fold up-regulated at 5 ℃ compared with that at 25 ℃,respectively.However,the expression levels of the two genes in 15 ℃ and 25 ℃ culture conditions showed no significant difference.Salicylic acid is one of plant hormones,which not only played an important role in the growth and development in plants,but also showed key roles in the biotic and abiotic stress resistance.In this macroalga U.prolifera cultured at 35 ℃,0.1 mmol/L salicylic acid promoted expression of MnSOD and CAT mRNA to 2.08 times and 5.30 times that of the control groups.But the expression levels of MnSOD and CAT mRNAs showed no significant difference from the controls when 0.01 mmol/L salicylic acid was added.In conclusion,the expression levels of MnSOD and CAT mRNAs enhanced in response to high or low temperatures.And the expression enhancement of the two genes was also one way of salicylic acid to alleviate the adverse effect caused by high temperature stress in U.prolifera.

    • Microbial carbon metabolic characteristics of biofilm communities in the grass carp culture pond based on Biolog-ECO plates

      2014, 38(12):1985-1995. DOI: 10.3724/SP.J.1231.2014.49435

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      Abstract:Microbial biofilm can be defined as multi-cellular aggregates adhering to carrier material,and has been widely used in water restoration.Despite prior research,the metabolic function of biofilm remains poorly understood.Using Eco-substrates as biofilm support,we evaluated the carbon requirements of microbial biofilm communities during biofilm formation(0,15,30,45,60 d)in the grass carp culture pond by Biolog-ECO plates.Our result showed that average well color development(AWCD)reached stationary phase after 168 h cultivation in all five selected sampling stages,and there were significant differences in sole carbon utilization characterized as AWCD value among five stages,and the higher carbon utilization capacity was observed in biofilm communities from 15 d to 45 d,it is higher than that observed in 0 and 60 d significantly.And the diversity indexes(Shannon index,Pielou index,McIntosh index,richness index)had the same rule of dynamic changes with AWCD.In addition,the polymers' and cabchydrates' utilization capacity of biofilm microbes was significantly higher than that of Phenolic compounds,carboxylic acids,amino acids,amines in all five sampling stages.With the biofilm formation,the utilization of α-D-glucose-1-phosphate,L-serine,N-acetyl-D-glucosamine,Tween 40,D-mannitol,etc.in biofilm microbes improved.The principal component analysis of microbial metabolic characteristics in biofilm communities showed that principal components 1(PC1)accounted for 33.9%,and principal components 2(PC2)accounted for 21.1% respectively.Based on samples' distance,most similar characters in carbon metabolism of microbial community were found among 15,30 and 45 d,but with significant differences in 0 and 60 d.Microbial biofilm communities had the strongest metabolic ability from 15 d to 45 d in the grass carp culture pond,and utilized carbon sources selectively.

    • Effects of air-exposure stress on antioxidant capacity and stress response of swimming crab(Portunus trituberculatus)

      2014, 38(12):1996-2004. DOI: 10.3724/SP.J.1231.2014.49441

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      Abstract:The purpose of this study was to evaluate the stress indicators which can reflect the physiology of swimming crabs(Portunus trituberculatus)sensitively during air-exposure and subsequent recovery.The effects of 0.5,1.5 and 3 h air-exposure stress on antioxidant capacity and stress response of crabs with an average weight of (87.93±1.24)g were investigated by determining hepatopancreas superoxide dismutases(SOD)activity,total antioxidant capacity(T-AOC),malondialdehyde(MDA)and heat shock protein 70(HSP70)level and muscles lactate contents.Crabs were sampled at 0.5,1.5 and 3 h during exposure to air,and at 0,2,4 and 10 h during following recovery,with crabs in seawater of (22.5±0.5)℃ as control.The main results showed that:(1)No significant effects of air exposure stress on SOD activity and MDA content in hepatopancreas were observed(P>0.05).However,air exposure stress had a significant influence on the T-AOC level,the content of HSP70 in hepatopancreas and muscles lactate contents.(2)After 0.5 h air-exposure stress,the SOD activity reached the maximum value when crabs had recovered for 4 h.During the recovery from 2 h to 10 h after 0.5 h stress,the MDA contents were significantly higher than that of control(P<0.05).No significant differences of the T-AOC level in hepatopancreas were observed during recovery after 0.5 h of air exposure(P>0.05).The SOD activity was similar to control after 1.5 h and 3 h of air exposure stress,and there were no significant differences between control and time intervals(P>0.05).The MDA content of crabs exposed to air for 1.5 h increased first and then decreased during recovery,with no significant differences observed between control and time intervals(P>0.05).However,after 3 h of air exposure,the MDA content significantly increased at 10 h during recovery(P<0.05).Besides,the T-AOC level reached the maximum value at 4 h after 1.5 h of air exposure,which was significantly higher than that of control(P<0.05).For crabs exposed to air for 3 h,the T-AOC significantly increased(P<0.05)at 0.5 h during recovery and then returned to the control level in 2-10 h(P<0.05).The lactate contents of all groups reached the maximum values at 0.5 h during recovery.The HSP70 levels of different treatments reached their maximum values in 2-4 h during recovery.These results indicated that,compared to other stress indicators which were selected,T-AOC level could more sensitively reflect the physiology of Portunus trituberculatus suffering air-exposure stress.SOD activities and MDA contents could be used synergistically to indicate the physiology of crabs during recovery.

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