Liaoning Ocean and Fishery Science Institute

Dalian, China

Liaoning Ocean and Fishery Science Institute

Dalian, China

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Shan H.W.,Ocean University of China | Bao W.Y.,Yangzhou University | Ma S.,Ocean University of China | Wei D.P.,Ocean University of China | Gao L.,Liaoning Ocean and Fishery Science Institute
Aquaculture International | Year: 2015

The toxicity of nitrogen in the shrimp culture water has been well established. In this study, SA beads composed of Vibrio alginolyticus VZ5, sodium alginate (SA) and sugarcane bagasse were used for ammonia nitrogen (NH4-N) and nitrite nitrogen (NO2-N) removal. A 50-day cultivation experiment was carried out in aquaria to evaluate the activity of the SA beads in shrimp culture. The results indicate that SA beads have a maximum capacity of 1.06 × 108 colony-forming units (cfu)/bead. However, the optimal initial density of the bacteria embedded in the SA beads is 104–105 cfu/bead. The maximum NO2-N degradation rate achieved for the SA beads was 8.44 mg/L/day, and the average NO2-N degradation per bead was 0.06 mg. The addition of a carbon source accelerated the degradation of NH4-N and NO2-N by the SA beads. The NH4-N and NO2-N concentrations after treatment with SA beads were below 1.55 and 1.62 mg/L, respectively, at later time points, and these concentrations were significantly lower than in the group without any treatment (P < 0.05, df = 17). There were no significant differences in the NH4-N and NO2-N concentrations following treatments with SA beads and water exchange (P > 0.05, df = 29), and the yield resulting from water treatment with SA beads reached approximately 70 % of the yield with water exchange treatment. Moreover, the particulate organic carbon and dissolved organic carbon concentrations in the water were enhanced by the addition of SA beads. At later time points, some of the SA beads had broken down, and the sugarcane bagasse from the SA beads may have served as a carbon source for forming bioflocs. The new approach proved effective for NH4-N and NO2-N removal in shrimp culture. © 2015 Springer International Publishing Switzerland


Ma Z.,Ocean University of China | Song X.,Ocean University of China | Wan R.,Ocean University of China | Gao L.,Ocean University of China | Gao L.,Liaoning Ocean and Fishery Science Institute
Ecological Indicators | Year: 2013

A modified water quality index (WQI) was used to evaluate the water quality of intensive culture ponds of Litopenaeus vannamei. Multivariate statistical technique, principal component analysis (PCA), was applied to interpret the large complex water quality data set generated over a span of 120 days (from 1st July to 28th October, 2008) with weekly monitoring of 11 variables in four different shrimp ponds. The PCA revealed three, four, three, and three latent factors that accounted for 82.843%, 81.6%, 81.629%, and 84.801% of the total variance in the water quality data sets from each of the four shrimp ponds, respectively. The varifactors indicated that the variables responsible for water quality deterioration were mainly related to organic matter group (COD and BOD 5), natural condition group (pH and T), and nutrient group (TAN, Chl-a and DIP) in intensive shrimp ponds. A modified WQI based on the varifactors was applied to evaluate the water quality in shrimp culture ponds. The result revealed that the overall water quality in the shrimp ponds were mainly excellent during the early period and deteriorated in the mid to late period. The WQI evaluates water quality synthetically; furthermore, it reveals the outcome when some of the other variables deteriorate significantly. Thus, this study illustrates the necessity and usefulness of multivariate statistical techniques for interpreting large and complex data sets regarding water quality in shrimp culture pond. Furthermore, the evaluation results revealed that the modified WQI can be used as a tool for determining water quality. © 2012 Elsevier Ltd. All rights reserved.


Gao X.-G.,Liaoning Ocean and Fishery Science Institute | Li H.-J.,Liaoning Ocean and Fishery Science Institute | Li Y.-F.,Liaoning Ocean and Fishery Science Institute | Sui L.-J.,Liaoning Normal University | And 4 more authors.
International Journal of Molecular Sciences | Year: 2010

The Chinese mitten crab (Eriocheir sinensis) is an economically important aquaculture species in China. In this study, we developed and evaluated simple sequence repeat markers from expressed sequence tags of E. sinensis. Among the 40 wild E. sinensis individuals tested, 16 loci were polymorphic. The number of alleles per locus ranged from two to ten. The observed heterozygosity ranged from 0.0667 to 0.9667, whereas the expected heterozygosity ranged from 0.0661 to 0.9051. These markers have the potential for use in genetic studies of population structure and intraspecific variation in E. sinensis. © 2010 by the authors; licensee MDPI, Basel, Switzerland.


Li H.,Liaoning Ocean and Fishery Science Institute | Zhu D.,Liaoning Normal University | Gao X.,Liaoning Ocean and Fishery Science Institute | Li Y.,Liaoning Ocean and Fishery Science Institute | And 2 more authors.
Conservation Genetics Resources | Year: 2010

In this study, we describe the first set of single nucleotide polymorphism (SNP) genotyping assays for hard clam (Meretrix meretrix) that were discovered through expressed sequence tag (EST) database mining. A total of 3,129 ESTs were assembled and 228 putative SNPs were identified from 51 SNP-containing contigs, with an average of 0.64 SNPs per 100 bp. A/G (T/C) and A/C (T/G) types had a high frequency and accounted for 46.2% and 35.6%, respectively. Thirty-two SNP genotyping assays were designed for validation based on allele-specific polymerase chain reaction (PCR) and melting temperature (T m)-shift analysis, and 15 (46.9%) of them showed polymorphisms with the minor allele frequency ranging from 0.075 to 0.475. Twelve (37.5%) assays failed due to amplification failure of one or both allele specific primers, and five (15.6%) assays lacked sufficient temperature discrimination for accurate genotype scoring. BLASTX gave significant hits for all 15 genotyped SNP-containing contigs but all were synonymous coding SNPs. These results indicate that EST data constitutes a valuable resource for SNP discovery and further maker development. © Springer Science+Business Media B.V. 2010.


Li H.-J.,Liaoning Ocean and Fishery Science Institute | Yang Q.,National Marine Environmental Monitoring Center | Gao X.-G.,Liaoning Ocean and Fishery Science Institute | Su H.,Liaoning Ocean and Fishery Science Institute | And 2 more authors.
Molecular Biology Reports | Year: 2012

LPS-induced TNF-α (LITAF) is a novel tran-scriptional factor that mediates the expression of inflammatory cytokines in LPS-induced processes. In the present study, the full-length cDNA encoding LITAF (designated as Mm-LITAF) was identified from Asiatic hard clam, Meretrix meretrix, by expressed sequence tag and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of Mm-LITAF was 1653 bp, consisting of a 5′ untranslated region (UTR) of 91 bp, a 3′UTR of 1166 bp with one cytokine RNA instability motif (ATTTA) and one polyadenylation signal (AATAAA), and an open reading frame (ORF) of 396 bp encoding a polypeptide of 131 amino acids with a theoretical isoelectric point of 7.49, and predicted molecular weight of 14.47 kDa. The deduced amino acid of Mm-LITAF shared 29-63% similarity with the LITAFs from other species, indicating that Mm-LITAF should be a member of the LITAF family. Two highly conserved CXXC motifs forming a compact Zn 2+-binding structure were also identified in Mm-LITAF. A quantitative reverse transcriptase real-time PCR (qRT-PCR) assay was developed to assess the expression of Mm-LITAF mRNA in different tissues, and the temporal expression of Mm-LITAF in clams challenged with Vibrio anguillarum. The mRNA transcript of Mm-LITAF could be detected in all the examined tissues with the highest expression level in the gill. Mm-LITAF expression was up-regulated significantly at 16 h in the gill and at 8 h in haemocytes after bacterial challenge, respectively. These results suggest that the Mm-LITAF is a constitutive and inducible acute-phase protein that perhaps involved in the innate immune response of hard clam. © 2011 Springer Science+Business Media B.V.


Li H.-J.,Liaoning Ocean and Fishery Science Institute | Liu W.-D.,Liaoning Ocean and Fishery Science Institute | Gao X.-G.,Liaoning Ocean and Fishery Science Institute | Zhu D.,Liaoning Normal University | And 3 more authors.
Molecular Biology Reports | Year: 2011

The hard clam Meretrix meretrix is an economically important shellfish in China. However, genomic research on this species is still at early stage, and few genomic resources are available. The objective of the present study was to generate expressed sequence tags (ESTs), and identify host-defense genes and microsatellite markers for M. meretrix. Three cDNA libraries for intestine, mantle and hepatopancreas were constructed using highly efficient SMART (Switching Mechanism At 5′ end of the RNA Transcript) method. A total of 3224 random clones were single-pass sequenced from 5′-ends, resulting in 3129 high-quality (>100 bp) ESTs averaging 734 bp. All the ESTs were assembled by software Cap 3, producing 1796 unigenes-1490 singletons and 306 contigs. All the unigenes were compared to the public protein database using tblastx, and 696 (38.8%) were homologues to known genes while the remaining 1100 (61.2%) appeared to be novel sequences. A total of 31 EST clusters were related to immune and defense functions. They included immune recognition receptors, proteases and protease inhibitors, and other immune-related genes. The screening of 1796 unigenes identified 55 (3.1%) microsatellite-containing sequences, with 20 having sufficient flanking sequences for primer design. Polymerase chain reaction amplification was successful for 12 primer pairs and 7 of them showed polymorphic. The EST collection and microsatellite markers obtained in this study provide a useful resource for further gene discovery and population genetic analysis in M. meretrix. © 2010 Springer Science+Business Media B.V.


Li H.,Liaoning Ocean and Fishery Science Institute | Jiang L.,Liaoning Ocean and Fishery Science Institute | Han J.,Liaoning Ocean and Fishery Science Institute | Su H.,Liaoning Ocean and Fishery Science Institute | And 2 more authors.
Fish Physiology and Biochemistry | Year: 2011

The major histocompatibility complex (MHC) is a large genomic region characterized by extremely high polymorphism and its association with resistance/susceptibility to disease in vertebrates. In this study, the full lengths of MHC IIA and IIB cDNA were obtained from spotted halibut (Verasper variegates) by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) approaches. The genomic structure, molecular polymorphism, and expression patterns were examined to study MHC II gene functions in fish. As in other teleosts, the genomic structure of the spotted halibut MHC IIA contained 4 exons and 3 introns. The deduced amino acid sequence of the class IIA molecule shared 28-79% similarity with those of teleosts and mammals. Nine class IIA alleles were identified from five individuals. Three alleles originating from a single individual suggested the existence of at least two class IIA loci in the genome. Six exons and 5 introns were identified from spotted halibut MHC IIB, and the deduced amino acid sequence shared 33-79% similarity with those of teleosts and mammals. Twelve alleles were identified, among which five were observed in a single individual, which suggested at least three class IIB loci. Quantitative real-time PCR analysis revealed the presence of class IIA and IIB transcripts in nine normal tissues with high expression level in kidney and gill. Furthermore, MHC IIA and IIB are probably two candidates of immune molecules involved in the acute-phase response in spotted halibut, because their transcriptional levels were significantly up-regulated in blood and liver after bacterial challenge. © 2011 Springer Science+Business Media B.V.


Yang Q.,National Marine Environmental Monitoring Center | Yang Z.,National Marine Environmental Monitoring Center | Li H.,National Marine Environmental Monitoring Center | Li H.,Liaoning Ocean and Fishery Science Institute
Fish and Shellfish Immunology | Year: 2011

Inhibitor of NF-κB (IκB) is an important member of Rel/NF-κB signaling pathway, which is an important mediator of immune responses in innate immune system. In this study, the IκB cDNA of hard clam Meretrix meretrix (designated as Mm-IκB) was cloned and characterized. The full-length cDNA of Mm-IκB was of 2098 bp, containing a 5′ untranslated region (UTR) of 123 bp, a 3′ UTR of 810 bp with a poly (A) tail, and an open reading frame (ORF) of 1164 bp encoding a polypeptide of 387 amino acids. The high similarity of Mm-IκB with other IκBs from invertebrates indicated that Mm-IκB should be a member of IκB family. Similar to most IκBs, Mm-IκB possessed all conserved features critical for the fundamental structure and function of IκBs, such as five ankyrin repeats and a conserved degradation motif (DS 44RYSS 48). Two PEST domains and a phosphorylation site motif (S 367EEE 370) at the C-terminus of Mm-IκB were identified. By quantitative real-time RT-PCR analysis, mRNA level of Mm-IκB was found to be most abundantly expressed in the tissues of mantle, gill and hepatopancreas, weakly expressed in muscle, foot and haemocyte. The Mm-IκB gene expression was significantly up-regulated at 24 h in haemocyte and at 12 h in gill after Vibrio anguillarum challenge, respectively. The results suggested the involvement of Mm-IκB in response against bacterial infection and further highlighted its functional importance in the immune system of M. meretrix. © 2011 Elsevier Ltd.


PubMed | Liaoning Ocean and Fishery Science Institute
Type: Journal Article | Journal: Molecular biology reports | Year: 2012

LPS-induced TNF- (LITAF) is a novel transcriptional factor that mediates the expression of inflammatory cytokines in LPS-induced processes. In the present study, the full-length cDNA encoding LITAF (designated as Mm-LITAF) was identified from Asiatic hard clam, Meretrix meretrix, by expressed sequence tag and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of Mm-LITAF was 1653 bp, consisting of a 5 untranslated region (UTR) of 91 bp, a 3UTR of 1166 bp with one cytokine RNA instability motif (ATTTA) and one polyadenylation signal (AATAAA), and an open reading frame (ORF) of 396 bp encoding a polypeptide of 131 amino acids with a theoretical isoelectric point of 7.49, and predicted molecular weight of 14.47 kDa. The deduced amino acid of Mm-LITAF shared 29-63% similarity with the LITAFs from other species, indicating that Mm-LITAF should be a member of the LITAF family. Two highly conserved CXXC motifs forming a compact Zn(2+)-binding structure were also identified in Mm-LITAF. A quantitative reverse transcriptase real-time PCR (qRT-PCR) assay was developed to assess the expression of Mm-LITAF mRNA in different tissues, and the temporal expression of Mm-LITAF in clams challenged with Vibrio anguillarum. The mRNA transcript of Mm-LITAF could be detected in all the examined tissues with the highest expression level in the gill. Mm-LITAF expression was up-regulated significantly at 16 h in the gill and at 8 h in haemocytes after bacterial challenge, respectively. These results suggest that the Mm-LITAF is a constitutive and inducible acute-phase protein that perhaps involved in the innate immune response of hard clam.


Li H.J.,Liaoning Ocean and Fishery Science Institute | He C.B.,Liaoning Ocean and Fishery Science Institute | Yang Q.,National Marine Environmental Monitoring Center | Shan Z.G.,Liaoning Ocean and Fishery Science Institute | And 2 more authors.
Aquatic Biology | Year: 2010

The Chinese mitten crab Eriocheir sinensis is an important aquaculture species in Asia and causes considerable economic and ecological damage as an invasive species in North America and Europe. Until recently, molecular markers available for genetic analysis in this species were limited to a few microsatellite sequences used in population studies. Here we describe the discovery of 3191 single nucleotide polymorphisms (SNPs) from the alignment of expressed sequence tags (ESTs) in E. sinensis. The observed frequency of SNPs was estimated at 0.78 per 100 bp of contig sequences. C/T substitutions were frequent and accounted for 32.2% of all SNPs. A subset of these SNPs (n = 38) was selected for validation by allele-specific PCR with melting temperature (Tm)-shift primers based on their observed frequency in sequence data; 12 (31.6%) of these were polymorphic in a panel of 40 wild-caught crabs. Eight (21.1%) did not amplify any product, and 18 (47.4%) failed due to amplification failure of 1 allele-specific primer. A table of optimal codons was deduced from the analysis of the E. sinensis EST dataset, and the results implied that the gene expression levels and GC-content might play important roles in codon usage bias. Sixteen codons ending with a G or C base were defined as 'optimal codons,' which may provide useful information for predicting gene expression in crabs. These are the first SNPs developed for the Chinese mitten crab and will provide a useful complement to currently available genetic markers. © Inter-Research 2010.

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