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Karyotype,Isozyme and Population Genetic Structure Analysis of Macrobrachium rosenbergii

ZHANG Lin, GAN Jinhua, ZHANG Tao, HE Li, ZHOU Jianguang

2023, 38(S1): 429-436. https://doi.org/10.7668/hbnxb.20193764

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In order to evaluate the germplasm resources of M.rosenbergii and provide reference for breeding guidance,it studied the chromosome karyotypes of M.rosenbergii using PHA,narcissin intramuscular injection and the sperms or ovaries cell direct production.The tissue expression patterns of lactate dehydrogenase(LDH),malate dehydrogenase(MDH),esterase(EST)and alcohol dehydrogenase(ADH)of muscle and gill of M.rosenbergii were analyzed by polyacrylamide gel electrophoresis(PAGE).The results showed that M.rosenbergii was diploid,2N=118=56M+24SM+10ST+28T,and arm number(NF)was 198.No sex chromosome or chromosome with satellite or secondary constriction was found.Four isozymes revealed 22 alleles and 15 genetic loci, and the effective number of alleles (Ne) were 1.47.Four enzymes had 7 polymorphic loci with gene frequencies ranging from 0.150 0 to 0.850 0.The proportion of polymorphic sites(P)was 46.7%.The average expected heterozygosity(He)value was 0.191 7,the average observed heterozygosity(Ho)value was 0.261 1,and the Hardy-Weinberg genetic deviation index(D)value was 0.379 7,indicating that there were both heterozygote excess and loss,but the heterozygote excess was more prominent,indicating that this population had a wide range of genetic variation.The degree of variation was large and the population genetic diversity was high,which could provide good parents for future artificial breeding.

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Genetic Diversity Analysis of Macrobrachium nipponense from Dalonghupao

LAN Ruolin, MA Wenzhi, ZHANG Limin, SUN Bo, CHANG Yumei, LIANG Liqun

2023, 38(S1): 437-443. https://doi.org/10.7668/hbnxb.20193703

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To learn more about the genetic characteristics of Macrobrachium nipponense and provide theoretical support for breeding selection,it used microsatellite molecular marking technology to compare the genetic diversity of Macrobrachium nipponense in Dalonghupao. The results showed that all 12 microsatellite loci were highly polymorphic loci (PIC>0.5). In the population of Taihu 2,the observed number of alleles ranged from 4 to 11,the effective number of alleles ranged from 3.686 4 to 7.480 5,the observed heterozygosity was 0.318 2—0.833 3,the expected heterozygosity was 0.744 9—0.884 8,the PIC of each locus was 0.691 9—0.851 7. The population of Taihu 2 was with high genetic diversity. In the population of Dalonghupao,the observed number of alleles ranged from 1—2,the effective number of alleles was 1.000 0—2.000 0,the observed heterozygosity was 0.000 0—1.000 0,the expected heterozygosity was 0.000 0—0.517 2,the PIC of each locus was 0.000 0—0.375 0. The population of Dalonghupao was with low genetic diversity. Seven loci in both populations deviated significantly from Hardy-Weinberg equilibrium,and three loci in the population of Dalonghupao could not be tested for Hardy-Weinberg equilibrium. Seven loci in the population of Taihu 2 showed heterozygous deletion,while seven loci in the population of Dalonghupao showed heterozygous excess. AMOVA analysis showed that 36.15% (P<0.01)of the total genetic variation was among populations and 63.85% (P<0.01)was among individuals. The genetic similarity between the population of Dalonghupao and the population of Taihu 2 was 0.276 9 and the genetic distance was 1.284 3,which showed that the genetic similarity between the two populations was low,and the genetic distance was far,and there were great genetic differences. The genetic differentiation index (F_ST)of the two populations was 0.361 53 (P<0.01),and the degree of genetic differentiation was obvious. The population genetic structure analysis showed an optimal K value of 3,indicating that the two populations had three possible ancestors. The level of genetic diversity of Dalonghupao wild population was low,which prompting us to protect the germplasm resources of the population of Dalonghupao.

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Electrophoretic Analysis on 3 Kinds of Isozymes in Gymnodiptychus dybowskii

ZHANG Tao, CHEN Peng, SHA Wenjun, WANG Ranxuan, HE Li, NIU Jiangong, GAN Jinhua

2023, 38(S1): 444-451. https://doi.org/10.7668/hbnxb.20193861

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This paper was to learn about the biochemical heredity characteristics and to enrich the research data about the germplasm resources of Gymnodiptychus dybowskii.Lactate dehydrogenase(LDH)isozymes,malate dehydrogenase(MDH)isozymes and Esterase(EST)isozymes in five kinds of tissues(heart,crystalline lens,liver,kidney and spleen)of Gymnodiptychus dybowskii were preliminarily studied by using the vertical polyacrylamide gel electrophoresis.The results showed that:LDH isozymes,MDH isozymes and EST isozymes showed tissue specificity and individual variations.The tissue specificity mainly reflected in expression of isozyme sites and degree of isozyme activity,which may be regulated by both heredity and metabolism and were related to the specific physiological functions of various kinds of tissues.LDH-C gene only expressed in liver.Its isozyme bands were near to negative pole in electrophoretogram, which were typical liver bands.In order to adapt the unique habitant,the isozymes of Gymnodiptychus dybowskii showed individual variations.Six supernate type of MDH isozyme bands expressed in heart may be the result of gene redundancy due to tetraploid during the evolution of Gymnodiptychus dybowskii.The research results could provide theoretical basis for the conservation and utilization of germolasm resources for Gymnodiptychus dybowskii from the biochemical genetics level.

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Electrophoretic Analysis on 3 Kinds of Isozymes in Schizothorax waltoni

ZHANG Tao, ZHANG Lin, GAN Jinhua, HE Li

2022, 37(S1): 366-372. https://doi.org/10.7668/hbnxb.20192824

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This paper was to learn about the biochemical heredity characteristics and to enrich the research data about the germplasm resources of Schizothorax waltoni.Lactate dehydrogenase(LDH)in five kinds of tissues(heart,crystalline lens,muscle,liver and serum),malate dehydrogenase(MDH)in five kinds of tissues(heart,crystalline lens,muscle,liver and kidney)and alcohol dehydrogenase(ADH)isozymes in liver of Schizothorax waltoni were preliminarily studied by using the vertical polyacrylamide gel electrophoresis.The results showed that:the band numbers of LDH in heart,crystalline lens,muscle,liver and serum were 6-10,8-10,4-7,0-7 and 1-7,respectively.The band numbers of MDH isozymes in heart,crystalline lens,muscle,liver and kidney were 6-7,2-6,5-7,5 and 3-5,respectively.There was 3 ADH bands in the livers.LDH-C gene only expressed in liver.LDH and MDH showed tissue specificity and individual variations.The tissue specificity mainly reflected in band numbers and degree of isozyme activity.There were correlations between tissue specificity and its physiological functions.In order to adapt the unique habitant,the isozymes of Schizothorax waltoni showed individual variations.

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Effects of Different Salinity,Alkalinity and pH Value on Sperm Motility and Fertilization Rate of Amur ide

LI Qian, LIANG Liqun, AI Keda, SUN Bo, ZHANG Limin, DONG Zhiguo, CHANG Yumei

2021, 36(S1): 381-389. https://doi.org/10.7668/hbnxb.20192551

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In order to determine the optimum range of salinity, alkalinity and pH value of sperm motility and fertilization rate of Amur ide. Different NaCl salinity(1‰, 2‰, 3‰, 4‰, 5‰, 6‰, 7‰, 8‰), NaHCO₃ alkalinity(10, 20, 30, 40, 50, 60, 70 mmol/L)and pH value(5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0)were set to stimulate the sperm motility of the Darinor Lake Amur ide.The sperm status of different treatment groups was observed, and the intense movement time, fast movement time and life span of the sperm were measured. Meanwhile, the fertilization rate in different salinity, alkalinity and pH solutions was determined.The results showed as follows. When the salinity ranged from 0 to 4‰, the sperm intense movement, fast movement time and life span increased gradually, and reached the maximum at 4. When the alkalinity was 0-50 mmol/L, the sperm intense movement, fast movement time and life span were prolonged with the increase of alkalinity, and reached the maximum at 50 mmol/L. In the pH value 5.0-7.0, the sperm intense movement, fast movement time and life span increased with the increase of pH, pH value 7.0 reached the maximum. The fertilization rates reached the highest when the salinity was 4‰, the basicity was 30 mmol/L and the pH value was 6.5, respectively. The fertilization rates in different salinity and alkalinity groups were significantly lower than those in the control group. In conclusion, based on the analysis results of different salinity, alkalinity and pH on sperm motility and fertilization rate of Amur ide, it was suggested to select saline-alkali water or fresh water with salinity of 0-4‰, alkalinity of 0-30 mmol/L and pH of 6.0-7.0 in the process of total artificial reproduction and cross breeding of Amur ide. It can obtain higher fertilization rate and seedling survival rate.

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Electrophoretic Analysis on Lactate Dehydrogenase(LDH)and Malate Dehydrogenase(MDH)Isozymes in Five Different Tissues in Schizothorax macropogon

ZHANG Tao, ZHANG Lin, ZHOU Jianguang, GAN Jinhua, HE Li

2021, 36(S1): 390-396. https://doi.org/10.7668/hbnxb.20192209

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This paper was to learn about the biochemical heredity characteristics and to enrich the research data about the germplasm resources of Schizothorax macropogon. Lactate dehydrogenase(LDH)and malate dehydrogenase(MDH)isozymes from five tissues(heart, crystalline lens, muscle, liver and kidney)of Schizothorax macropogon were preliminarily studied by using the vertical polyacrylamide gel electrophoresis. The results showed that:the band numbers of LDH isozymes expressed in heart was 6-7, in crystalline lens 6-10, in muscle 5-7, in liver 8-11 and in kidney, 8-10 respectively, and the band numbers of MDH isozymes expressed in heart was 4-8, in crystalline lens 2-4, in muscle 3-4, in liver 4-5 and in kidney 4-7. LDH-C gene only specificly expressed in liver. The band numbers expressed in liver are the largest in five different tissues. MDH bands expressed in liver presented the darkest and showed the most degree of isozyme activity. LDH isozymes and MDH isozymes showed tissue specificity and individual variations. The tissue specificity mainly reflected in band numbers and degree of isozyme activity. There were correlations between tissue specificity and its specificly physiological functions. In order to adapt the unique habitant, the isozymes of Schizothorax macropogon showed individual variations. The research results can provide theoretical basis for making germplasm standards and the conservation and utilization for Schizothorax macropogon from the biochemical genetics level.

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Analysis of Water Quality and Bacterial Diversity of Aquaculture Water of Tilapia

ZHU Lifei, LI Wei, ZHOU Yuqi, LIU Huijun, ZHANG Xin, WANG Saisai, SUN Yansheng

2020, 35(S1): 416-423. https://doi.org/10.7668/hbnxb.20191229

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To investigate the effect of tilapia cultivation density on water quality parameters and bacterial diversity,water quality monitoring was carried out. After 30 days of breeding experiment,water samples were collected to detected the diversity and abundance of bacteria though Illumina Miseq 16S rDNA gene sequence,meanwhile histopathological examination was conducted. The results showed that the rearing density of tilapia did not affect liver tissue. With the increase of rearing density,gills presented pathological changes such as twist,enlarged ends and even loss of branchial lobule. The dissolved oxygen content in aquaculture water decreased with the increase of rearing density(P<0.05). The correlation coefficient matrix of water quality parameters and load analysis showed that the nitrate nitrogen and nitrite nitrogen in the water of different culture density groups were moderately correlated. Dissolved oxygen in the low-density and high-density groups was negatively correlated with ammonia nitrogen in medium intensity,while dissolved oxygen in the medium-density group was negatively correlated with ammonia nitrogen. The rearing densities lead to different abundance of bacteria in aquaculture water. The dominant bacteria were Flavobacterium, Sphingomonas, hgcl_clade,and Macrosomonas and so on,and most of them had the ability to degrade pollutants. The above results showed that adequate oxygen supply in medium-culture density of tilapia could promote the degradation of inorganic nitrogen in water,and the combination of effective water treatment could ensure the growth of fish,which could bring the maximum economic effect.

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Cloning and Expression Analysis of JHEH Gene in Portunus trituberculatus

ZHENG Liang, XIE Xi, ZHENG Hongkun, XU Dongjie, ZHU Dongfa

2020, 35(6): 225-234. https://doi.org/10.7668/hbnxb.20191281

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To explore the role of JHEH in crustaceans, the full-length cDNA of JHEH from the swimming crab, Portunus trituberculatus (PtJHEH) was cloned and characterized using the RACE technique followed by bioinformatics analysis, and its gene expression levels were detected in different tissues, molting cycles and ovarian development stages using the Real-time quantitative PCR(RT-qPCR). The obtained PtJHEH was 1 803 bp in length, containing an open reading frame(ORF) of 1 374 bp, which encoded 467 amino acids. The deduced amino acid sequence was predicted to have transmembrane domains, signal peptides, and typical epoxide catalytic domain. The qPCR results showed that PtJHEH was expressed extensively in various tissues, but the highest mRNA level was found in the hepatopancreas, which was significantly higher than other tissues(P<0.05). This might be the reflection that the hepatopancreas was the main site of crustacean metabolism. During the molt cycle, the PtJHEH in the hepatopancreas expressed at highest level in the pre-molt D2 stage, while its expression in Y-organs was remarkably high at the post-molt stages(A and B stages), but then decreased significantly. During the ovarian development process, the expression of PtJHEH in hepatopancreas and Y-organs both increased to the highest level at the late-vitellogenic stage(stage Ⅲ). The expression pattern of PtJHEH in the molting cycle and ovarian development showed a correlation to the fluctuation in the hemolymph MF titers previously reported by our group. This indicates that JHEH may have a role in the regulation of MF degradation.

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Analysis of Codon Usage Bias in Carassius auratus

ZHANG Xin, YOU Hongzheng, LI Rongni, ZHU Lifei, LI Wentong, LIANG Yongjun, SHI Dongjie, SUN Yansheng

2017, 32(S1): 30-36. https://doi.org/10.7668/hbnxb.2017.S1.006

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The study of codon usage bias in species can be in-depth understanding of gene expressions,and provide the basis for molecular breeding.8 143 eligible coding DNA sequences were selecting from transcriptome data of Carassius auratus which were obtained through transcriptome sequencing by Illumina HiSeq 2000.CDS were analyzed by CodonW and CUSP programs in C.auratus genome,calculating frequency of codon usage which encoded amino acids and comparing with variety classes of mode organisms such as:Homo sapiens,yeast, E. coli, Drosophila, D.rerio. The consequences indicated that twenty-eight optimal codons of C.auratus were identified in 64 codons,effective number of codons was 54.9,GC content of genes was 0.509,GC content at the third position of synonymous codon was 0.564 the codons which were ending with G or C were bias using.And codon bias of C.auratus was difference in various degrees with other mode organisms,codon usage frequency of C.auratus was biggest different from yeast,but had a consistency with codon bias of D.rerio.

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Cloning, Expression and Activity Analysis of Two Kinds ACADs in Cyprinus carpio

YU Wenjuan, YU Juhua, LI Hongxia, LI Jianlin, TANG Yongkai, YU Fan, HE Feng, FU Chunjie

2017, 32(3): 48-57. https://doi.org/10.7668/hbnxb.2017.03.008

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To investigate the regulatory mechanism of fatty acids fatty Acyl coenzyme A (Acyl-CoA dehydrogenase,ACADs) on fatty acids β-oxidation in mitochondrial,this experiment cloned two genes,MCAD and LCAD,of ACADs family from the liver tissue of Cyprinus carpio using classic RT-PCR.The open reading frames of MCAD and LCAD were 1 275,1 326 bp in length respectively,encoding 424 and 441 amino acids with characteristics of ACADs family structure (such as FAO binding sites,the substrate binding site and catalytic site),sharing high homology with zebrafish,93.16% and 94.56% respectively.The results of RT-PCR showed that MCAD and LCAD mainly expressed in the liver and heart. No significant influence was observed of different sources of fat on the expression of MCAD,while fish oil could obviously stimulate the expression of LCAD. After IPTG induction,both the two prokaryotic expression vectors,MCADs-pEASY(E2),and LCADs-pEASY(E2),expressed fusion proteins of 43.0,43.6 kDa in molecular weight,with obvious absorption peak at 370,450 nm,which was proved to be able to naturally bind to FAO cofactors. Phenazine methyl sulfate reaction method was applied to determine the enzyme activity of Prokaryotic expression protein,and the results indicated that the optimum temperature for MCAD and LCAD enzymes were 28℃,and the enzyme activity were 9.12,10.29 U/g respectively. This study preliminary confirmed that MCAD and LCAD had highly expression in liver and heart of Cyprinus carpio,similar as in Mammals,and high unsaturated fatty acid could promote the expression of LCAD.

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Polypeptide Analysis of GIFT Tilapia Liver after Infecting Streptococcus agalactiae

ZHU Jiajie, FU Qiang, AO Qiuwei, CHEN Ming, TAN Yun, LI Liping, JIANG Hesheng, GAN Xi

2016, 31(S1): 485-489. https://doi.org/10.7668/hbnxb.2016.S1.083

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The polypeptide expression on GIFT tilapia liver was studied after infecting Streptococcus agalactiae.Matrix-assisted Laser Desorption Ionization Time of Flight (MALDI-TOF) was employed to analyze the polypeptide,and they were identified by mass spectrometry.The results indicated that there were 44 and 52 polypeptide between control group and experimental group,which Signal/Noise (s/n) was more than 50.We discovered 6 peaks (m/z 1 082, 1 161,1 196,1 459,1 740,1 877) was obvious difference in two groups(P<0.01).The 1 196(m/z) peptide decreased,but the 1 161 and 1 459(m/z) peptide increased in experimental group.All the mass spectrums were corresponding to 19 peptides,and 14 shows a up-regulated expression,4 shows down-regulated expression,and 1 shows no obvious change.The results of bioinformatical analysis indicated that all the peptides were involved in three regulation pathways including TCA cycle,DNA splice and cell defensive signaling pathway.Real-time PCR was utilized to analysis the mRNA variation of ATP synthase 6 (ATP6) gene and D-beta-hydroxybutyrate dehydrogenase (Bhd1) gene.The results confirmed the positive correlation of gene expression and protein expression.The results of peptidome analysis provided a new approach to diagnose the Streptococcus agalactiae disease.

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Expression Characterization of Interferon Regulatory Factor 1 in Nile Tilapia

WANG Dan, XIE Wenjie, ZHU Yefei, ZHAO Jinliang, CHEN Xiaowu

2016, 31(4): 31-38. https://doi.org/10.7668/hbnxb.2016.04.006

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In order to study the expression and function of interferon Regulatory Factor (IRF1) in nile tilapia,the full length cDNA of IRF1 was cloned and applied for phylogenetic analysis,and the expression variation was checked by qRT-PCR after PolyI:C stimulation.Cell transfection by eukaryotic expression vector was used to analyzing for subcellular localization.The opening reading frame of nile tilapia IRF1 contains 888 bp with deduced protein of 295 amino acids.Structurally,nile tilapia IRF1 protein typically shares the conserved characterizations with other species' IRF1 homologues,displaying conserved DNA-binding domain,IRF association domain,serine-rich C terminal domain,and 6 tryptophan residues in the N terminal.The results of phylogenetic analysis showed that the IRF family was divided into four subfamily:IRF1,IRF3,IRF4 and IRF5.The nile tilapia IRF1 was found to be merged into the teleost subgroup,amphibian and mammalian homologues were clustered into their corresponding subgroups.Real-time PCR revealed a broad expression pattern of nile tilapia IRF1 in various tissues.The expression of IRF1 was mainly in the head kidney,spleen,liver and gill,furthermore,which increased obviously after PolyI:C stimulation,especially for the spleen and head kidney.Subcellular localization analysis showed that nile tilapia IRF1 mainly resides in the cytoplasm.These data supported the view that nile tilapia IRF1 was a potential molecule in IFN immune defense system against viral infection.

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Application of Microsatellite Markers in Endangered Hucho taimen Enhancement and Releasing

TONG Guang-xiang, KUANG You-yi, ZHANG Yong-quan, YIN Jia-sheng

2015, 30(S1): 38-45. https://doi.org/10.7668/hbnxb.2015.S1.007

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In order to study the application of microsatellites markers in endangered Hucho taimen enhancement and releasing hucho releasing,eighteen microsatellites markers were used to analyze the genetic diversity and structure of release-recapture population (JC),Tahe River population (TH) and released parents' population (HT,YZ) in the present study.The recaptured individuals were also identified.The results indicated that Na,Ne,I,He and H indexes exhibited the same tendency,HT > JC > TH > YZ.The observed heterozygosity (Ho) was highest in YZ population while lowest in TH population.All of these four populations possessed a medium level of genetic diversity.AMOVA analysis showed that there was no significant difference within four groups while extremely significant difference between populations.The genetic similarity was highest between YZ and HT population with the index of 0.718 3,medium-level between JC population and other three populations with the index of 0.566 5-0.689 5, and lowest between YZ and TH population with the index of 0.495 9.The results showed that we could understand the genetic background of the parents and assure the released descendant's polymorphism by microsatellite markers.The released individuals were identified by microsatellite markers.In conclusion,this study provided the basis for the next stage of artificial releasing work.

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The Influence of Replacing Fish Meal with Soybean Meal on Growth,Body Composition and Digestive Ability of Jian Carp in Practical Extruded Diets

JIANG Jun, ZHOU Xiao-qiu, YIN Long, FENG Lin, JIANG Wei-dan, LIU Yang, ZHAO Ye

2015, 30(S1): 107-111. https://doi.org/10.7668/hbnxb.2015.S1.020

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The present study was performed to investigate the effect of replacing fish meal with soybean meal on growth,body composition and digestive ability of Jian carp.A total of 720 Jian carp with average initial weight of(240.2±5.7) g were randomly allocated four groups of each three replicates.Four isonitrogenous and isoenergetic practical extruded diets were formulated by replacing 0,25%,50%,and 100% fish meal with soybean meal.The results showed that there was no significant difference in the percent weight gain(PWG) and the specific growth rate(SGR) in fish fed diets by replacing 0,25%,and 50% fish meal with soybean meal.(P> 0.05).Fish fed the diet by replacing 100% fish meal with soybean meal had significantly lower PWG and SGR than the 0 group(P<0.05).The crude protein in whole body were significantly lower when the replacement ratio of fish meal by soybean meal was 100%(P<0.05).Fish fed the diet by replacing 100% fish meal with soybean meal had significantly lower,γ-GT and AKP than the 0 group(P<0.05).These results indicated there was no significant difference on growth performance of juvenile Jian carp at body weight 200-800 g when soybean meal could be used by replacing 50% of fish meal in practical extruded diets.

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Genetic Diversity of Leuciscus waleckii Populations Based on CO ⅠSequence

DOU Xin-jie, CHANG Yu-mei, TANG Ran, SUN Xiao-wen, TAO Ran, WANG Nan, LIANG Li-qun, ZHANG Li-xia

2014, 29(S1): 29-34. https://doi.org/10.7668/hbnxb.2014.S1.007

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To analyzing the genetic diversity and genetic structure of Leuciscus waleckii based on mtDNA CO Ⅰgene(635 bp length),six geographical populations(72 samples) were sampled from Dali Lake(DL),Ganggeng Lake(GG),Hulun Lake(HL),Amur River(HLJ),Songhua River(SHJ) and Wusuli River(WSL).On the base composition,A+T content(52.37%) was slightly higher than that of G+C content(47.63%),corresponding to the characteristics of nucleotide composition in the vertebrates;On the genetic parameters,the genetic diversities of Dali Basin populations(DL and GG) were lower than those of Amur Basin counterparts(HL,HLJ,SHJ and WSL);There were 20 haplotypes,of one haplotype was shared by all the six populations,one shared by Dali Basin and one shared by Amur Basin.Despite there were significant population differentiation between Dali Basin and Amur Basin(P<0.05) as well as genetic structure,the analysis of AMOVA and the values of frequent gene exchange(Nm) both demonstrated that the six Leuciscus waleckii populations had very close relationship,probably derived from a common ancestor,and had not yet differentiated to different species or subspecies based on present evidence.

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Raising Dali lake Leuciscus Waleckii (Dybowski) in Saline-alkali Pond

LI Jia, ZHANG Feng-qin, Sun Cai-xia, ZHANG Yong-wang, ZHAO Xiao-bing, SHI Li-dong, LIAN Rui-jun, LIANG Wen-gung, BAN Shun-li, MA Xiao-tang

2006, 21(S3): 76-78. https://doi.org/10.7668/hbnxb.2006.S3.015

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To test the raising technology and productivity of raising Leuciscus waleckii (Dybowski) in 2814" saline-alkali pond in Huhhot, 150000 larvae from Dali lake were taken to the pond. After 4 months feeding, growth rate of Dybowski in this saline-alkali pond was 2 to 3 times faster than Dali lake. The results showed that Dybowski was a good fish category for pond raising of Inner Mongolia Autonomous region.

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The Survey of Fibrin Fit Content in Fish Feedstuff

CUI Zhi-feng, HAO Yan-zhou, SUN Wei-ming, LI Deng-lai

2006, 21(S3): 79-81. https://doi.org/10.7668/hbnxb.2006.S3.016

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Baidu(2)

Crude fiber (CF)is a important nutrient for fishes.The roles of CF, optimum content in the compounded diet of fishes and effective factors, and harm of excessive content of CF are summarized.The methods of research are also evaluated.The future studies are proposed.

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Test on Hatch of Amur Ide(Leuciscus Waleckii(Dybowski) Egg in Different Kind of Water in Dali Lake

MENG He-ping, PENG Ben-chu, WANG Bao-wen, ZHANG Li, WANG Zhi, HAN Guo-cang, GAO Yu-kui, LIU Yu-hai

2006, 21(S3): 152-154. https://doi.org/10.7668/hbnxb.2006.S3.035

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Baidu(1)

We done experiment to hatch Amur ide egg in different kind of water and experiment to survival Amur ide fry in Dali lake, Keshiketeng banner, Chifeng city, the result of these experiment showed that the Amur ide egg can be hatched and the Amur ide fry can survive better in Qigainaoer lake, Erdos city. The result of these experiment set up a example for transfer Amur ide from Dali lake to part region of Inner Mongolia.

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Effect of Different Temperature Inducement on Growth and Development of the Larvae Genetic Improvement of Farmed Tilapia

ZHU Jia-jie, LIN Yong, CHEN Zhong, LI Li-ping, TANG Zhan-yang, YANG Hui-zan, LUO Yong-ju, GAN Xi

2010, 25(S2): 170-173. https://doi.org/10.7668/hbnxb.2010.S2.038

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Baidu(4)

"In this research,the effects of different temperature inducement on the growth rate,survival rate and aberration rate of the larvae of Genetic Improvement of Farmed Tilapia(GIFT)was studied in the laboratory.The results showed that the growth rate and survival rate of the larvae were significant different after 10 days inducement,but the aberration rate was not obvious.The growth rate and survival rate of the larvae were highest at 28℃,the lowest was at 38℃.The mortality rate at 39℃was 100% after 10 days inducement.Moreover,the growth rate at 28℃ was significant different to the other temperature.Above-mentioned results indicated that high temperature plays very important role in the larvae of the growth rate and survival rate.So we should pay attention to regulate the temperature of the process of the larvae GIFT cultivated,in order to improve the quality of tilapia offspring. "

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Semi-quantitative RT-PCR Method of Detection of Heat Stress on Fish Liver Content of HSP70 mRNA

SU Ling, LI Shao-wu, WANG Di, LIU Hong-bai, LU Tong-yan, YIN Jia-sheng

2010, 25(S1): 100-104. https://doi.org/10.7668/hbnxb.2010.S1.023

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Baidu(9)

This research investigated the variation of HSP70 mRNA content in the liver of Carassius auratus at different time lapses after heat stress processing by using semiquantitative RT-PCR. Two primers were designed for this study according to the sequences of Carassius auratus's heat shock protein70(HSP70) and beta-actin mRNA published by GenBank. The results show that the HSP70 expression in liver tissue is time-dependent by showing an increase-to-decline trend. The expression starts to increase 1 hour after inducing and reaches a maximum 4 h later.Subsequently,it begins to decline continuously and eventually returns to its initial value until 48 h later.

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Identification of Microsatellite Markers Associated with Growth Rate Traits in Hucho taimen

TONG Guang-xiang, KUANG You-yi, ZHANG Chao, YIN Jia-sheng

https://doi.org/10.3969/j.issn.1000-7091.2012.z1.007

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22 microsatellite markers were used to analyze the growth rate related traits of 176 individuals fromF1 of random mating Hucho taimen， including body weight， body length， body height， body width， head length， headwidth and interorbital width． The results showed that of the 67 alleles detected in total， the numbers of alleles rangedfrom 2 to 6 per locus with the fragments of 109 to 452 bp， and the mean observed heteozygosity and the mean expected heteozygosity were 0． 583 5 and 0． 564 0， respectively． Deviations from Hardy-Weinberg equilibrium ( HWE)occurred at 22 loci in different degrees． The normal distribution of 7 traits was analyzed using SPSS software and theresults indicated that all the seven traits had a characteristic of continuous variation belonging to the typical quantitative traits or polygenic inheritance traits， and conformed to the normal distribution． Meanwhile， GLM linear modelwas constructed between the 22 microsatellite markers and the growth rate related traits， and the markers significantly correlative to the traits were performed multiple comparison based on the genotypes． The results revealed that of22 microsatellite loci， there were 11 markers significantly correlative to one trait at least， and the minimum numberof markers related to body length was 4 and the maximum number of markers related to body height and body widthwas 7． Through the multiple comparisons， it was found that there was significant difference of genotypes from thesame trait in every locus， and there was also difference among the mean value of the different markers． The screeningof these markers could provide some basis for the further study on molecular marker-assisted breeding．

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Isolation and Characterization of Microsatellite from Genome of Aspiorhynchus laticeps

HAO Zhuo-ran, LIANG Li-qun, CHANG Yu-mei, REN Bo

https://doi.org/10.3969/j.issn.1000-7091.2012.z1.008

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Magnetic beads enriched method was used to isolate microsatellite DNA from Aspiorhynchus laticepsgenome． Aspiorhynchus laticeps genomic DNA was extracted and then digested with restriction enzyme Sau3 AI． Targeted segments of 250 － 750 bp were collected by centrifugation of sucrose density gradient and ligated to adaptors ．Then the purified ligated DNA was hybridized with Biotin-labeled simple sequence repeats probes( CA) 8，( TGA) 8．After capture of target fragments by magnetic beads and PCR amplification， the selected DNA s were cloned into thepMD18 － T vector and transformed into competent Escherichia coli DH5α， and finally a microsatellite library wasobtained． 563 positive colonies were obtain through twice screens by colony PCR． Sequencing of these positive colonies confirmed that 53 contained microsatellite loci( number of repeats ≥ 5 ) ． In these sequences， 36 repeat motifs( about 67． 92% ) were perfect， 12 repeat motifs ( about 22． 64% ) were imperfect， and 5 repeat motifs ( about9. 43 % ) were compound． We design 24 pairs of primers from 50 microsatellite sequence and compose them． As aresult， 56 pairs were screened and used successfully to amplify special fragment， among which 24 pairs were polymorphism． Using 24 pairs of these microsatellite markers， we analyzed genetic structures of one Aspiorhynchus laticeps population． The result showed that the alleles were ranged from 3 － 6， while the polymorphic information content( PIC ) were 0． 625 3 ． Twelve of the loci were in high polymorphic level．

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Effects of the Chinese Herbal Compounds on the Heat Shock Protein 70 Expression of Crucian Carp

LI Shao-wu, WANG Di, SU Ling, LIU Hong-bai

2012, 27(S1): 401-405. https://doi.org/10.3969/j.issn.1000-7091.2012.z1.078

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In this study， real time quantitative PCR method was performed to understand the effects of fourkinds of Chinese herbal compounds， including AH， CD， Ⅳ and APS on the expression of heat shock protein 70 inthe liver and brain tissues of Crucian carp． A total of 60 fishes were divided into 5 groups and fed on diet with 3 %Chinese herbal compounds of body weight． After feeding for 30 days continuously， the expression of HSP 70 in theliver and brain tissues was investigated both under the optimum temperature ( 18℃ ) and heat shock temperature(28℃ ) for an hour． The results indicated that the expression of HSP 70 in control group increased in varying degreeduring the heat shock stress， and it was significantly higher in the liver than that of brain tissues both under the normal condition or heat shock stress． Of four experimental groups， the content of HSP 70 mRNA was found to increasein AH， IV and APS groups while decrease in CD group after heat shock stress， significantly decreasing in the livertissues． In addition， AH Chinese herb， with the main function of replenishing blood and Qi， could greatly enhancethe expression of HSP 70 in the liver of Crucian carp by 4． 86 folds and 3 ． 43 folds compared to control group under18℃ and 28℃ ， respectively． In conclusion， AH compound was found to promote high-level expression of HSP 70gene in Crucian carp， to be helpful to resist heat shock and could be applied as an effective anti-heat stress drug．

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Genetic Diversity of Populations in Germplasm Resources of Houttuynia cordata Thunb

ZHONG Jun, WANG Kun, QIU Ping

https://doi.org/10.7668/hbnxb.2009.S2.019

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The test wanted to post variation law and genetic diversity by studying different regional populations of Houttuynia cordata Thunb germplasm resources in Huaihua city of Hunan province, and supplied farther evidences for protection and utilization of resources. The differentiation coefficients and genetic parameter and diversity index analysis was applied on the population phenotypic traits of 16 germplasm resources of Houttuynia cordata Thunb from Huaihua city of Hunan Provinces. The frequency is different in the three quality traits which green overground stem, white underground stem and cordiform leaf were representative phenotypic quality traits. The variation coefficients of branch number and fresh weightiness of overground stem, the node number of underground stem, 4-methyl-1-cypropyl-3-cyclohexoally-1-alcoholy and 2, 4-methyl-3-cyclohexoallic cypropyl alcoholy between populations were the higher, whereas the variation coefficient of root diameter was the lowest ;but those of undecanone and branch number of overground stem per plant were lowest(13.4%)and highest respectively(39.8%)on the phenotypic traits within populations. The phenotypic diversity index between populations(0. 213 -0. 804)was relatively higher than that of within populations(0. 182 -0.710), the lowest in root number and the highest in underground fresh weightness ;the phenotypic diversity index of different geographic population was 0. 574 -1. 024 with the highest in Huangyan prefecture and the lowest in Jigongjie prefecture. The mean variation coefficients of between population accounted 1/2 whereas that of within population was 1/3 which the variations between populations were main variation source. The largest phenotypic differentiation coefficient between populations was the branch number of overground (77.04 %)whereas the smallest was root diameter(51.88 %). The absolute genetic advances were smaller (RGS% < 40%) but the heritabilitys were larger(h~2 > 50%)to the most quantity traits under 5% selection intensity. Cluster analysis based on morphological traits showed that the populations could be divided into two and three groups in genetic distance 2.0 and 1.5 respectively Which cluster in same kind was according as firstly color of overground stem, leaf shape next, altitude heigh again, region last. The differentiation coefficients were rich and the differentiation degree were high in populations.

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Species Identification of Cytb Gene in Fish Product

ZHANG Hong-yi, CHAI Ping-hai, MEI Yan-qiao, DENG Tian-ning

2011, 26(S2): 9-12. https://doi.org/10.7668/hbnxb.2011.S2.003

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In this study Theragra chalcogramma,Hippoglossus hippoglossus,Merlangius merlangus and Limanda limanda were identified by sequence method,based on Cytb gene.The preliminary results showed that ITS1 was similar with Theragra chalcogramma(DQ356946) by 99%,ITS2 was similar with Hippoglossus hippoglossus(EU492151)by 98%,ITS3 with Merlangius merlangus(AY946299) by 98%,ITS4 with Limanda limanda(EU492119) by 99%.We also detected 50 samples about fish authenticity identification,and testing results also showed that most samples are consistent with their name.

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Optimization of Isolation and Primary Culture of Hepatocytes in Crucian Carp (Carassius auratus)

JIA Rui, CAO Li-ping, DING Wei-dong, DU Jin-liang, XU Pao, YIN Guo-jun

2011, 26(S2): 206-212. https://doi.org/10.7668/hbnxb.2011.S2.044

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In order to establish a stable primary culture model of hepatocytes in Carassius auratus,3 different isolation methods(tissue culture,mechanical speration and pancreatin digestion) and culture conditions of primary hepatocytes culture were compared in the present study.The survival rate was tested with trypan blue exclusion and the viability was assessed with WST-1;the morphological changes of cells in long-term culture were also observed.The results showed that in tissue culture,hepatocytes migrated from liver and proliferated after 4-5 days,compare with the other methods,it took much longer time for the hepatocytes to move out and it was difficult to collect the cells for further culture.The number and viability of the hepatocytes obtained by mechanical separation were obviously lower than by pancreatin digestion.The liver tissue digested by 0.1% trypsin for 30 min yielded 1.47×108 cell/g(liver weight) and the viability was 91.43%.Both culture medium and concentration of serum influenced the attachment and proliferation of the hepatocytes.The cell cultivated in M199 basal medium supplemented with 10% fetal calf serum at 28℃,4% CO2 for 8-9 days can sub-cultured

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Effects of Recombination HSP70 on Immune Function of Tilapia Peripheral Blood Lymphocyte

CHEN Ming, WANG Qiu-hua, WANG Rui, GAN Xi, LI Li-ping, LEI Ai-ying, LIANG Wan-wen, HUANG Wei-yi

2011, 26(S2): 213-218. https://doi.org/10.7668/hbnxb.2011.S2.045

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In this report,we use the recombinant tilapia Hsp70(tHsp70)from Pichia yeast to prepared non-covalent HSP70-antigen complexes with thallus and extracellular bacteria products(ECP)of Streptococcus iniae,and investigated their effects on cellular immune response of tilapia by detection of Proliferation and related immune gene expression of peripheral blood lymphocyte in vitro.Our results demonstrated that tHsp70 could form complexes with thallus and ECP of tilaplia in vitro.In addition,tHsp70(10 μg/mL)and all tHsp70(10 μg/mL)-thallus complexes enhanced the proliferation of tilapia peripheral blood lymphocytes(P0.01).Similarly,thallus alone also enhanced the proliferation of tilapia peripheral blood lymphocytes(P0.05).By contrast,ECP alone exhibited some inhibitory effect on the proliferation of tilapia peripheral blood lymphocytes(P=0.056).However,the complex of tHsp70 at 10,but not at 100 and 1 μg/mL,with ECP reduced the inhibitory effect of ECP on the proliferation of tilapia peripheral blood lymphocytes.Moreover,tHsp70(10 μg/mL)alone or tHsp70(10 μg/mL)-antigen complexes could significantly influence the expression of immune related genes(PGK,MMP9,ICER,MHCⅡ)in tilapia peripheral blood lymphocyte compare to antigen alone and medium control groups.The study indicated that tHsp70 have the immunoadjuvant and the immunopotentiator function,and provided experimental data for development the tHsp70 based vaccine against tilapia streptococcal diseases.

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Influence of Two Kinds of Microecologics on the Digestive Enzymes in Grass Carp

WANG Di, LI Shao-wu, YIN Jia-sheng, LU Tong-yan

2011, 26(S2): 219-223. https://doi.org/10.7668/hbnxb.2011.S2.046

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Two kinds of microecologics(named M-Ⅰ and M-Ⅱ),which were based on the Bacillus subtilis and had already been developed and applied in the practice,were selected to determine their influence on several digestive enzymes in grass carp.M-Ⅰ was used to mix with the feeding and M-Ⅱ was poured into the water of the ponds.Four groups were set in this study,including the control group,M-Ⅰ group,M-Ⅱ group,and the combination group of these two microecologics.After the usage of microecologics for 3 months,the amylase,protease and lipase of the livers and intestines were determined and compared with each other.The results indicated that M-II preparations,which mainly functioned on regulating the water quality,did not affect the three digestive enzymes in grass carp,while M-I preparations,which could promote digestion and growth of fishes,were found to greatly improve the enzyme activities.In addition,the mix usage of M-Ⅰ and M-Ⅱ were proved to play a role in both regulating the water quality and promoting digestion,which would further promote the growth and reproduction of grass carp.Therefore,it could be concluded that microecologics were benefit for the rapid and healthy growth of grass carp farmed in the ponds.

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Separation and Cultivation of Tilapia Peritoneal Macrophage

WANG Qiu-hua, CHEN Ming, HUANG Wei-yi, WANG Rui, LI Li-ping, GAN Xi, LEI Ai-ying, HUANG Jun, LIANG Wan-wen

2011, 26(S2): 224-228. https://doi.org/10.7668/hbnxb.2011.S2.047

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In order to study factors that influence the separation and culture of tilapia peritoneal macrophages,and to recognition the characteristic of tilapia peritoneal macrophages.In the study,We separated macrophages from peritoneal cavity of Tialpia(Oreochromis niloticus♀×Oreochromis aureus♂)after intraperitoneal injected with squalene in the condition of difference growth temperature;The cell characteristic was observed by screening cell medium and Wright's staining.The results show the macrophages were easily obtained in the condition of 19~25℃,Separation result was obviously affected when water temperature was aboved 28℃;It was a optimization period for separation between 48-72 h after injected squalene;Serum of tilapia play an important role in the tilapia peritoneal macrophages invitro cultrue and fetal bovine serum was not fit;The characteristic of tilapia peritoneal macrophage was similar to mammalian macrophage,irregularly shaped with a low nucleus to cytoplasm ratio,adherent to plastic and glass,able to form multi-nucleated giant.The work laid a foundation for the further research of immunological function about tilapia peritoneal macrophage.

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Isolation and Purification of Lipovitellin from Thymallus arcticus grubei and Preparation of Anti-serum

SUO Li, QU Qiu-zhi, SUN Da-jiang, ZHANG Ying, WANG Xue, ZHAO Ji-wei

2011, 26(S2): 229-234. https://doi.org/10.7668/hbnxb.2011.S2.048

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The yolk protein is the main nutrients during embryonic development.The quantity and quality of yolk is crucial for early larval growth.We have purified the egg curde extract of Amur grayling by Sephadex G-200.The results showed there were three protein peaks 1,2 and 3 by gel chromatographv at 4℃ 0.2 mmol/L PMSF and three protein peaks were relatively symmetric.Then protein peak 2 was purified by more than one time Sephadex G-200 eluting.The properties of yolk protein were analyzed by the methods of polyacrylamide gel electrophoresis and immuno diffusion.Based on the 7.5%Native-PAGE staining data,three brands were detected with molecular weights of 567.2,486.3,157.5 kDa.Based on the 12%SDS-PAGE staining data,nine brands were detected with molecular weights of 80.7,68.7,46.8,39,30,20.9,16.7,14.9,14.1 kDa.Protein peak 2 was lipovitellin and molecular weight was 486.3 kDa.Lipovitellin was composed of three subunits with molecular weights of 80.7,30,14.9,14.1 kDa,respectively.Polyclonal antibodies against the purified Lv from Amur grayling were produced in the New Zealand rabbit.Double immunodiffusion showed that the determination of the titre for anti-Lv serum was 1 ∶ 64.Double immunodiffusion showed that anti-Lv serum have sex-specificity.

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Compaire Study of Biology on Two Breeding Aquatics Models in Paddyeield of Crayfish

XU Zeng-hong, ZHAO Chao-yang, ZHOU Xin

2011, 26(S1): 248-251. https://doi.org/10.7668/hbnxb.2011.S1.050

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Compaire two different models about paddyfiled breeding aquatics of crayfish,i.e.the model A,putting parents crayfish to paddyfield,increase and breeding unartificial,live throught the winter natural.the model B,setting up breeding canopies in paddyfield for reproducing and throught winter warmly.From Oct.to the next Mar.,mensurating the series of indexes including spawning time and rate,baby crayfish growing speed,the size and rate at the end of harvest time,the breeding cost and so on,to evaluate two breeding models effect.The results show that the model B is better than the model A evident,the time of the parents crayfish spawn move up,the spawning rate heighten,the growing speed of baby crayfish quicken,sampling stat.at the harvest time in jun.,in model A,the rate above 30 g is 30.8%,20-30 g is 45.5%,below 20 g is 23.7%,in model B,the rate above 30 g is 52.6%,20~30 g is 30.9%,below 20 g is 16.5%,it indicates that the model B outgoes the model A.The study also compaire the effects of the different feeds sorts,the small fish mixed manpower feedstuff or the small fish singleness are better than the manpower feedstuff singleness.So study the different breeding models of crayfish to confirm to put in how many the reasonable young number,breeding entironment and feeding modes,the study can to provide practice experiences and science gist for breed aquatics cosmically of crayfish.

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Effects of Dietary Vitamin D_3 Supplementation on Body Composition and Activity of Alkaline Phosphatase in the Serum of Juvenile Mirror Carp(C. carpio Songpu mirror carp)

ZHANG Tong, XU Qi-you, XU Hong, WANG Chang-an, SHI Lian-yu

2011, 26(S1): 258-263. https://doi.org/10.7668/hbnxb.2011.S1.052

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This experiment was conducted to study the effect of vitamin D3 levels on body composition,concentration of calcium and phosphorus and activity of alkaline phosphatase(ALP) in the serum of juvenile mirror carp(C.carpio Songpu mirror carp.),to determine the best dietary vitamin D3 level of juvenile carp.525 juvenile mirror carps,weighting an average of 2.4 g were randomly assigned to seven dietary treatments with three replicates and 25 carps with each pen.Seven levels of vitamin D3(0,200,400,800,1 600,3 200 and 1×105 IU/kg) of the dietary were fed to seven groups.The feeding trial lasted for 56 days.The results showed that moisture,crude protein,crude lipid contents of the body and concentration of phosphorus in the serum were not significantly affected by vitamin D3(P0.05).The crude ash,calcium and phosphorus contents of the body,concentration of calcium and activity of ALP in the serum were significantly affected by vitamin D3(P0.05),compared to the experimental group which without vitamin D3.Calcium and phosphorus contents of the body,concentration of calcium and phosphorus and activity of ALP in the serum were increased and then decreased with increasing of dietary vitamin D3 supplementation.These results indicated that the calcium and phosphorus contents of the body,concentration of calcium and activity of ALP in the serum was significantly increased by suitable vitamin D3 level,compared with no or excessive vitamin D3 level.The suitable requirement of juvenile mirror carp is about 708.3-769.8 IU/kg diet,based on establishing indexes line model to calculate the calcium,phosphorus contents of the body and ALP activity of mirror carp in the serum.

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The Correlation and Path Analysis for Growth-related Traits of GIFT Strain Tilapia(Oreochromis niloticus)

YANG Hui-zan, LIN Yong, TANG Zhang-sheng, LI Jun, GAN Xi

2011, 26(S1): 264-268. https://doi.org/10.7668/hbnxb.2011.S1.053

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In order to study the effects of morphometric characters on body weight in GIFT Strain Tilapia(Oreochromis niloticus),545 one year old individuals(female:290,male:255)were sampled for measuring the metric traits,including the body length(x₁),head length(x₂),trunk length(x₃),body depth(x₄),body width(x₅) and body weight(y).

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Study on the Growth of Hucho taimenJuvenile at Different Temperatures

WANG Jin-yan, ZHANG Ying, YIN Jia-sheng

2011, 26(S1): 274-277. https://doi.org/10.7668/hbnxb.2011.S1.055

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The growth characteristics of Hucho taimen juveniles were studied at different temperatures by aquarium.The results showed that the instantaneous growth rate was significantly affected by the different water temperatures.The most suitable tempearture range was 15-18℃,and upper temperatures was 24℃.As the results also express as curve regression,the optimum model of body length growth was Quadratic(R20.98),and that of the body weight growth was Cubic(R20.98).The growth model was competent to estimate the culture production.The artificial feed can be used to rear the fish as the main food,and the condition factor of fish fed on formulated feed was close to that of fish fed on Limnodrilus and Copepoda.

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Analysis of Cold Hardy Capacity in Eight Strains Tilapia and Their Hybridizations

LIN Yong, LU Qi-xi, YANG Hui-zan, CHEN Zhong, HUANG Yin, GAN Xi, ZENG Lan, TANG Zhang-sheng

2011, 26(S1): 278-282. https://doi.org/10.7668/hbnxb.2011.S1.056

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Compared the cold hardy capacity in different tilapia strains for providing cold hardy capacity tilapia breeding with breeding materials and theoretical support.Through the indoor artificially low-temperature stress,measured eight strains tilapia and 15 hybridized by tilapia varieties′ cold-tolerance,and compared their cold hardy capacity.In 3 strains aureus tilapia Oreochromis aureus,the semi-lethal low temperature respectively were:the USA strains 7.82℃,summer strains 8.22℃,guangdong strains 8.44℃;and mortality respectively were:56.76%,62.25%,79.43%;in 5 strains Nile tilapia the semi-lethal low temperature respectively were;the USA strain 8.86℃、Egypt strain 9.39℃,Guangdong strain 9.30℃,Thailand strain 9.57℃,Gift strain 10.13℃,and the mortality respectivelywere:70.43%,56.43%,67.24%,83.54%,91.66%;In 15 hybrid strains,the strongest cold-tolerance was the strain from Nile tilapia of Egypt strain as female and the USA strain of aureus tilapia Oreochromis aureus as paternal,that semi-lethal low temperature was 8.22℃ and mortality was 51.06%.And semi-lethal low temperature and mortality of correlation showed significant between each species and varieties.Hybrid strains tilapia′ cold hardy capacity were highly correlated with their parents,their cold-tolerance may inheritance by cytoplasmic.To strengthen the screening of the female can obtain faster genetic progress.Through cold hardy capacity performance comparison,determinated the hybridization tilapia from the Egypt strain of Nile tilapia as female and the USA strain of aureus tilapia Oreochromis aureus as paternal for cold hardy capacity breeding materials.

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Screen of Disease}esistance Molecular Markers and Analysis of Genetic Diversity in GIFT Strain Nile Tilapia

ZHU Jia-jie, ZHOU Yu, SHEN Xia-shuang, LI Li, LUO Yong-ju, GAN Xi

2013, 28(S1): 16-21. https://doi.org/10.7668/hbnxb.2013.S1.004

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Technology with microsatellite was used to analyze the genetic difference between disease-resistant and disease-susceptible groups of GIFT strain. The results show that there were 20 loci successfully amplified with stable target bands within 30 tested loci to identify the polymorphism for the tested GIFT strain. The average alleles in disease-resistant was 2. 5 slightly higher than that of 2. 4 in disease-rusceptible group. Then the average of observed heterozygosity(Ho) and expected heterozygosity(He) were 0. 469 and 0. 441, 0. 440 and 0. 476 respectively and the polymorphism information content (PIC) was 0. 359 and 0. 379. Genetic similarity index and genetic distance was 0. 055 and 0. 054 respectively. The locus UNH845,UNH189 and GM180 showed significant departure from Hardy Weinberg equilibrium after sequential bonferroni correction(P<0. 05)in two groups. GM251 and GM462 sites differences of allele fragment(135,198 bp) were positively related to disease-resistance GIFT strain. Above mentioned results indicated that the genetic diversity between disease-resistant and disease}usceptible group of GIFT strain was not significantly different, and the GM251 and GM462 gene fragment with its differential bands from the PCR ampliftcation can be used as the molecular marker for the disease}esistant GIFT strain molecular assistant selection.

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Microsatellite Marker's Isolation，Primer Design and Application in the Goldfish

ZHANG Sheng-li, YANG Pu, LIANG Yong-jun, SUN Xiang-jun, SUN Yan-sheng, SU Jian-tong, SHI Dong-jie

2013, 28(S1): 27-31. https://doi.org/10.7668/hbnxb.2013.S1.006

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Microsatellite DNA was isolated from goldfish with combining biotin capture method. We obtained 38 positive clones and isolated 31 microsatellite sequences. 12 pairs of primers were designed by Primer Premier 5.0 and 8 pairs of primers can obtain amplifying products among them. The genetic diversity of the artificial breed goldfish populations were studied by using the 8 Microsatellite primers. The results showed that the genetic diversity level of two populations were high. The average heterozygosity(Ho) in two populations was from 0. 534 3 to 0. 565 5,average excepted heterozygosity(He) was from 0. 628 6 to 0. 657 0,and average polymorphism information content (PIC) was from 0. 500 9 to 0. 510 6. The genetic distance was 0. 095 7 and genetic identity was 0.921 1.All of 8 microsatellite locus could be used for genetic analysis in goldfish.

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Study on the Genetic Diversity of the Conservation Population in Taimen

ZHANG Chao, TONG Guang-xiang, KUANG You-yi, YIN Jia-sheng

2013, 28(S1): 113-119. https://doi.org/10.7668/hbnxb.2013.S1.022

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The genetic diversity of the taimen conservation population(Contained 4 groups) with 29 polymorphic microsatellite loci. The study showed that in the 332 sampled individuals,2一13 alleles were detected in each locus,the average effective allele number was 3. 961 3,the average observed heterozygosity and expected heterozygosity were 0. 642 and 0. 675 1 respectively, the average PIC was 0. 633.The genetic diversity of the 4 groups was B7(F1♀× wild ♂3×3) exceed B10(F1♀×F1♂3×3),B10(F1♀×F1♂3×3) exceed B6(wild♀×wild♂3×3),B9(F1♀×F1♂1×1)was the lowest. Except the group F1♀×F1♂1×1,the genetic distance between B6(wild♀×wild♂3×3)、B9(F1♀×F1♂1×1)、 was the farthest between B7(F1♀×wild♂3×3) and B10(F1♀×F1♂3×3))with the nearest distance. The clustering result based on the genetic distance showed that B6(wild♀×wild♂3×3),B9(F1♀×F1♂1×1),B10(F1♀×F1♂3×3)clustered as one Glade respectively,the cross breeding groups B7(F1♀×wild♂3×3)was divided into two Glades. This research also discussed the possibility that the endangered animals to maintain the rich diversity in offsprings.

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Comparison on Salt Tolerance and Growth between Family Offsprings and Mixed Group of Oreochromis niloticus ♀ × Sarotherodon melanotheron ♂ F₂

WANG Fei, JIA Jin-hua, ZHANG Yan-hong, REN Bing-chen, ZHAO Li-hui, ZHUANG Qing-qing, LIANG Cong-fei, YAN Biao, ZHAO Jin-liang

2013, 28(S1): 278-282. https://doi.org/10.7668/hbnxb.2013.S1.052

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F₂ family offsprings and mixed group were obtained by paired and mixed mating the hybrid F, individuals of Oreochromas ualotacus × Sarotherodou melauotherou a,respectively. The salt tolerance and growth under different salinities ( 0,15,20,25 g/L) of juvenile fishes from two mating modes were compared. The results showed that MST, STS,MLS6 of 0. uiloticus×S. melauotherou a Fz were higher than that of the control group 0. ualotacus(P<0.05).No significant differences in MST, STS。 and MLS}6 indexes were observed between the family offsprings and mixed group ( P>0.05)except for the family 2. Under the same salinity conditions,there warn’t significant difffernces in the average growth rate between the family offsprings and mixed group (P>0.05) with a exception of family 5 in 0 g/L senility, uiloticus × S. melauotherou a Fz exhibited the lowest weight toefficient of variation at 25 g/L senility. The results provided some basic data for hybrization and utilization of 0.uiloticus × S. melauotherou.

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Study on the Effect of Body and Gonad Growth of Crayfish Feeding Biology Feed

XU Zeng-hong, ZHOU Xin, SHUI Yan, ZHAO Chao-yang

2013, 28(S1): 283-286. https://doi.org/10.7668/hbnxb.2013.S1.053

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Compare the effect of feeding biology feed and general feed to breed crayfish to value the biology feed effect by observing and mensurating the crayfish’s growth rate and the gonad growth. The results show that the biology feed are excelled than the general feed on the amino acids kinds and the content of the EAA especially namely the otherness on EAAI is prominent. The average growth rate of the biology feed and general feed are 1. 75 % and 1 .48% after two months feeding. There are some different degree otherness on the gonad growth coeffidents , the ratio of the liven to body and the comparative eggs quantity, after math statistic checkout, the otherness on body growth and gonad growth between biology feed and general feed to feed caryfish is prominent, the study indicate that the biology feed have good value to crayfish and aquiculture.

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Population Characteristics of Amur Ide Leuciscus Waleckii (Dybowski)in Dali lake

MENG He-ping, ZHANG Li, WANG Bao-wen, ZHANG Jiang-hua, SONG Zhi-gang, PENG Ben-chu, DU Zhao-hong, HAN Guo-cang, GAO Yu-kui, LIU Yu-hai

2007, 22(S3): 124-127. https://doi.org/10.7668/hbnxb.2007.S3.031

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Through measure,analysis on the morphological characteristics and the part biological characteristics of Amur ide`eaptured during its reproductive&grow reason,this paper showed that Amur population age structure in Dali lake is unreasonal,there are some questions exist in Dali lake fishery;such as fisheries resource decline and over-capture,so the capture size of Amur ide should be decided that the body length is more than 17cm,the body weigh is more than 83g.Base on the results of these work,we draw up the regression equations between body length, body weigh and age.We found out that the Alkaline and PH is so high and the Cat is very scarce that the fishery is limited in Dali Iake.

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Population Characteristics of Golden (chinese) Carp Carassius Auratus (Linnaeus) in Dali Lake

ZHANG Li, MENG He-ping, LI Zhi-ming, PENG Ben-chu, YUAN Man-chuan, DU Zhao-hong, AN Ming, HAN Guo-cang, GAO Yu-kui, LIU Yu-hai

2007, 22(S3): 128-130. https://doi.org/10.7668/hbnxb.2007.S3.032

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Through measure,analysis and study on the morphological characteristics and the part biological characteristics,this paper showed that Carassius auratus grow style in Dali lake is homogeneous,the population reproductivity is better,but the population age structure is unreasonal,so the number of C}u-assius auratus卿 breed by man should be increased and the capture number in Dali Lake should be decreased.The reasonable capture size for Carassius auratus in Dali lake is that the body weight is more than 214.Sg,the body length is more than 19.67cm.

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Examination and Analysis on the Infection by Aeromona of Fish and Pathogeny

CHEN Cui zhen, ZHANG Xiao jun, FANG Hai, HE Zhen pingWANG Xiu yun, GONG Yuan fang

2000, 15(S1): 178-184. https://doi.org/10.3321/j.issn:1000-7091.2000.Z1.038

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The paper examined the conditions of diseases and pathological changes of fish to the natural infection of euteritis of grass carp, putting a sed of silver carp, rotting gill of grass carp, albinism of silver carp, haemorrhage of carp body surface infect and septicaemia of soft-shelled turtle infect. In the meantime, pathogeny was examined by using the materials of pathological changes. The result showed that all the infection were caused by Aeromonas. Through identifiying80 strains of isloated bacteria, the result showed A. hydrophila was 29 strains (36.25 per cent), A. sobria was 13 strains (16.25 per cent), A. caviae had38 strains (47.5 per cent), all of the diseases iufeeted by single or more pathogewic bacteria.

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The Effects of Rare Earth Elements on Yield of Commen Carp

JIANG Zhen-ying

2005, 20(S1): 122-124. https://doi.org/10.7668/hbnxb.2005.S1.031

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Based on feed rare earth elements to commen carp,The effect rare earth elements on the production Increment of commen carp are studied.The experimental results that the rare earth elements could raise the yield 14.1%(9.3%~16.8%)and increase the economic results 8335.6yuan/hm².

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Orthogonal Design and Optimization of SRAP Amplification System for Macrobrachium nipponense

YAO Jian-hua, FU Hong-tuo, GONG Yong-sheng, WU Yan, HE Xin-long, LI Hui-ping, QIAO Hui

2008, 23(S2): 122-125. https://doi.org/10.7668/hbnxb.2008.S2.027

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Four factors ( Taq DNA polymerase,dNTP,Mg²⁺, primer) with four levels were studied by orthogonal design in order to establish SRAP technique of Macrobrachium nipponense. Firstly, effective amplification systems were screened out, which were optimized at the second step. Opt imal amplif icat ion system for Macrobrachium nipponense wasestablished with 2. 5 mmol/ L Taq DNA polymerase, 0.??25 mmol/ L dNTP, 0.64 U/ 20L Mg2+ and 0.6 mol/ L primer.The result laid on foundation for the application of SRAP marker in Macrobrachium nipponense.

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Composition of Pathogenic Bacteria in Litopenaeus vannarnei Culture Ecosystem

ZHANG Hong-yi, ZHAO Yong, DAI Xi-lin, PAN Ying-jie

2008, 23(S1): 257-262. https://doi.org/10.7668/hbnxb.2008.S1.060

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Vibrio is one of the most dominant pathogens. Using 16 S rDNA clone library method, the Vibrio composiion of the Litopenaeus vannarnei Culture Ecosystem was investigated. Compared with the published sequences in Gen-Bank data base, sequencing results of cloned 16 S rDNA amplicons revealed a diverse community in the Litopenaeus vannarnei Culture Ecosystem. The results indicated that Water Sample was clustered into 16 OTUs, the dominant commun-ities are Vibrio, Chlorof lexi and Pantoea, which accounted for 33.3%, 18.5% and 7.41%; sediment samplewas clusteredinto 14 OTUs, the dominant communities are Vibrio and Pantoea, which accounted for 50.0% and 13.6%, respectively.The results had a little difference from the two samples: there were not Aeromonas in the Water Sample andthere were not proteobacterium in the sediment sample.

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Exogenous Xylanase Influences the Activities and Distribution of Six Digestive Enzymes in Gaster, Intestine and Hepatopancreas of Tilapia nilotica L.

2006, 21(4): 123-130. https://doi.org/10.3321/j.issn:1000-7091.2006.04.031

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Wheat basal diet was control.The tested diets were wheat basal diet added with different level of xylanase(0.05%,0.10%,and 0.15%).Each treatment was devised with 5 repeats and each repeat had 40 male Tilapia nilotica L.All the fishes lived in floating cages were fed 4 times a day and each time were fed to satiation.75 days later the activities of proteinase,amylase,lipase,xylanase,cellulose and pectinase were determined.The results indicated that:The activities of proteinase and cellulose in gaster of the 0.10% experimental group increased significantly compared with those of the control(P<0.01).The activities of the six digestive enzymes in intestine of all experimental groups increased in different levels.Especially,the activities of all the six digestive enzymes in intestine of the 0.10% experimental group increased significantly(P<0.01).Exogenous xylanase inhibited the activities of proteinase,amylase and lipase in hepatopancreas,but promoted the activities of endogenic xylanase,cellulose and pectinase.Adding exogenous xylanase did not effect on the distribution of amylase,endogenic xylanase,cellulose and pectinase in gaster,intestine and hepatopancreas,but had distinct influences on the distribution of proteinase and lipase.

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1981, 1(3): 79-88. https://doi.org/10.7668/hbnxb.1981.03.023

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1979年我们和石家庄地区的岗南水库、横山岭水库、唐山市的陡河水库合作进行网箱养鱼的生产性试验。试验用网箱均为单架浮动式网箱。各水库的网箱规格有所不同:岗南水库的长7米，宽4米，高2米，横山岭水库的长5米，宽5米，高2米，陡河水库的长8米，宽4米，高2米，此外尚有其他一些规格的网箱。网箱材料均由乙纶线编织，网目自1.1—1.5公分不等。试验以养殖鲢、鳙鱼鱼种为主，也有少量试验成鱼。网箱养殖的鱼种不投喂人工饲料，依赖库水中的天然饵料——浮游生物。

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Preliminary study on Phytoplankton and Potential Fishery Productivity of Silver and Bighead Carps in Intensive Cultivated Carp Ponds

Liu Guocai

1994, 9(3): 122-126. https://doi.org/10.3321/j.issn:1000-7091.1994.03.025

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The biomass of phytoplankton in intensive cultivated carp ponds in Tongliao was between 23.6-53.8mg/ L.Scenedesmus,Chlorococcum,Chlorella,Coelastrum etc,which were contained in Chlorococcales were more in there ponds,and the amount of these species in Chlorococcales was 43% in that of the total biomass of phytoplankton.Cryptomonas,Euglena etc,which had flagellum also had certain amount,and was 37% in that of the total phytoplankton biomass.The primary productivity was 402.6kg(wet weight)/ h•m²•day,and it could provide 1553.4kg/h•m²•day output of the silver and bighead carp.

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Itestinal Contents of Wintering Fishes and Their Absorbing Nourishment Under Ice

Liu Guocai, Wen Haishen, Cui Yajie

1996, 11(3): 128-132. https://doi.org/10.3321/j.issn:1000-7091.1996.03.025

Abstract (336) PDF (215) RichHTML

Under low temperature of winter season,wintering fishes can absorb nourishment from intestinal contents(organic pieces,algae,water plants,silts).The saturation and composition of intestinal contents change with different types of water and fishes.The saturation in the intestine is increased gradually from front intestine,and the latter part of it has the highest saturation and can reach fourth degree.The research result showed that feeding wintering fishes could insure and increase survival rate and growth.

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