Lingbing Zeng

Chinese Academy of Fishery Sciences, 北江, Zhejiang Sheng, China

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Publications (20)34.09 Total impact

  • Nan Jiang · Jin Xu · Jie Ma · Yuding Fan · Yong Zhou · Wenzhi Liu · Lingbing Zeng
    Wuhan University Journal of Natural Sciences 10/2015; 20(5). DOI:10.1007/s11859-015-1114-9
  • Yong Zhou · Yuding Fan · Scott E LaPatra · Jie Ma · Jin Xu · Yan Meng · Nan Jiang · Lingbing Zeng
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    ABSTRACT: The major capsid protein (MCP) is the main immunogenic protein of iridoviruses, that has been widely used as an immunogen in vaccination trials. In this study, the codon-optimized giant salamander iridovirus (GSIV) MCP gene (O-MCP) was synthesized and cloned into a pPICZα B vector for secretory expression in the methylotrophic yeast Pichia pastoris after methanol induction. The expression of the O-MCP protein was detected by the Bradford protein assay, SDS-PAGE, Western blotting and electron microscopy. The Bradford protein assay indicated that the concentration of the O-MCP expressed was about 40μg/ml in culture supernatants. SDS-PAGE analysis revealed that the O-MCP had a molecular weight of about 66kDa and reacted with a His-specific MAb that was confirmed by Western blotting. Electron microscopy observations revealed that the purified O-MCP could self-assemble into virus-like particles. Healthy giant salamanders were vaccinated by intramuscular injection with the O-MCP antigen at a dose of 20μg/individual. The numbers of erythrocytes and leukocytes in the peripheral blood of immunized Chinese giant salamanders increased significantly at day 3 and reached a peak at day 5 post-immunization. Meanwhile, the differential leukocyte counts of monocytes and neutrophils increased significantly at day 5 post-immunization compared to that of the control group. The percentage of lymphocytes was 71.33±3.57% at day 21 post-immunization. The neutralization assay showed that the serum neutralizing antibody titer reached 321 at day 21 post-immunization. The GSIV challenge test revealed that the relative percent survival of Chinese giant salamanders vaccinated with O-MCP was 78%. These results indicated that the O-MCP antigen expressed by the Pichia pastoris system elicited significant immune response in the Chinese giant salamander against GSIV and might represent a potential yeast-derived vaccine candidate that could be used for the control of disease caused by the giant salamander iridovirus. Copyright © 2015. Published by Elsevier Ltd.
    Vaccine 08/2015; DOI:10.1016/j.vaccine.2015.08.054 · 3.62 Impact Factor
  • Nan Jiang · Yuding Fan · Yong Zhou · Wenzhi Liu · Jie Ma · Yan Meng · Congxin Xie · Lingbing Zeng
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    ABSTRACT: The Chinese giant salamander iridovirus (GSIV), belonging to the genus Ranavirus in the family Iridoviridae, causes severe hemorrhagic lesions and nearly 100% mortality in naturally infected Chinese giant salamanders Andrias davidiamus. However, the replication and distribution of the virus has not been well characterized in vivo. Using in situ hybridization, the expression of the GSIV major capsid protein (MCP) was detected in the cytoplasm of cells of the spleen, kidney, liver and gut tissues. MCP expression in the spleen and kidney appeared to fluctuate significantly during the acute phase of infection. Using an immunofluorescence assay, GSIV antigens were abundant in the spleen and kidney tissues but appeared to be at relatively low levels in the liver and gut. Additionally, there were significant changes in the expression of the pro-inflammatory cytokines macrophage migration inhibitory factor (MIF), tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β) in different tissues in response to infection with GSIV. The expression of MIF, TNF-α and IL-1β had significantly increased in the spleen at 3 d post-infection; this correlated with a decrease in virus replication in the spleen. These results suggest that the spleen and kidney are the major target tissues of GSIV, and the increased expression of MIF, TNF‑α and IL-1β may contribute to a reduction of virus replication in the spleen.
    Diseases of Aquatic Organisms 06/2015; 114(3). DOI:10.3354/dao02868 · 1.75 Impact Factor
  • Jie Ma · Nan Jiang · Scott E. LaPatra · Ling Jin · Jin Xu · Yuding Fan · Yong Zhou · Lingbing Zeng
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    ABSTRACT: Haematopoietic necrosis of gibel carp (Carassius auratus gibelio) is caused by cyprinid herpesvirus 2 (CyHV-2) and has caused huge economic losses in aquaculture worldwide. Currently the isolation and propagation of CyHV-2 in vitro is very difficult due to the lack of permissive cell lines. Studies on the pathogenesis of CyHV-2 have been hampered because the virus has not been extensively characterized. In this study, a novel cell line from the brain of gibel carp, denoted GiCB, has been established and characterized. Sustainable propagation of CyHV-2 in the GiCB cell line has been confirmed by virus infection and titration, PCR, transmission electron microscopy, immunofluorescence assay and fluorescence in situ hybridization. The GiCB cells showed typical cytopathic effect by day 6 post-infection with CyHV-2 including cell shrinkage, rounding, and cell fusion with cytoplasmic vacuolization. The virus titer reached 10(7.5±0.37)TCID50/ml and has been successfully passaged over 50 times in the GiCB cell line. Electron microscopy analysis revealed the complete replication of CyHV-2 in GiCB cells. CyHV-2-infected GiCB cells reacted strongly with polyclonal antibodies against CyHV-2 and CyHV-2 RNA in cells hybridized specifically with the virus RNA probes. Additionally, an experimental infection demonstrated that CyHV-2 produced in GiCB cells caused 100% mortality in gibel carp. All the results provide solid evidence that the GiCB cell line is highly permissive for the isolation and propagation of CyHV-2. This is a significant advancement that will promote additional research on CyHV-2 infection in fish in the future. Copyright © 2015 Elsevier B.V. All rights reserved.
    Veterinary Microbiology 04/2015; 177(3-4). DOI:10.1016/j.vetmic.2015.04.006 · 2.51 Impact Factor
  • Jin Xu · Xianhui Zeng · Nan Jiang · Yong Zhou · Lingbing Zeng
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    ABSTRACT: Chinese sturgeon (Acipenser sinensis) is a rare and endangered species, and also an important resource for the sturgeon aquaculture industry in China. Recently, a fatal bacterial disease occurred in farmed Chinese sturgeon. The clinical signs of the diseased fish were systemic blood loss and anemia. The gills were very pale with swollen and hemorrhagic gill filaments. Discrete blood spots were apparent in the oral cavity. Small petechial hemorrhages were present in pale livers and the digestive tract. Hemorrhages were also present in the kidney and gonad. The spleen was red and swollen. Additionally, a small amount of clear ascites fluid was present in the body cavity. A bacterial pathogen was isolated from the internal organs of diseased fish and subsequently identified as Pseudomonas alcaligenes by morphology, biochemical testing and phylogenetic analysis of the 16S rDNA. Experimental infection in healthy hybrid sturgeon confirmed the pathogenicity of the bacterial isolate. The results of this study represent the first isolation and identification of Pseudomonas alcaligenes in cultured Chinese sturgeon in China and provide a foundation for the diagnosis, prevention and treatment of this new disease.
    Aquaculture 04/2015; 446. DOI:10.1016/j.aquaculture.2015.04.014 · 1.88 Impact Factor
  • Qian Chen · Jie Ma · Yuding Fan · Yan Meng · Jin Xu · Yong Zhou · Wenzhi Liu · Xianhui Zeng · Lingbing Zeng
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    ABSTRACT: The type I IFNs play a major role in the first line of defense against virus infections. In this study, the type I IFN gene designated gsIFN was identified and characterized in the Chinese giant salamander (Andrias davidianus). The genomic DNA of gsIFN contains 5 exons and 4 introns and has a total length of 5622bp. The full-length cDNA sequence of gsIFN is 1113bp and encodes a putative protein of 186 amino acids that has a 43% identity to type I IFN of Xenopus tropicalis. The deduced amino acid sequence has the C-terminal CAWE motif, that is mostly conserved in the higher vertebrate type I IFNs. Real-time fluorescence quantitative RT-PCR analysis revealed broad expression of gsIFN in vivo and the highest level expression in blood, kidney and spleen. Additionally, the expression of gsIFN at the mRNA level was significantly induced in peripheral blood leucocytes after stimulation with poly I:C and after infection with the Chinese giant salamander iridovirus (GSIV). A plasmid expressing gsIFN was constructed and transfected into the Chinese giant salamander muscle cell line. Expression of the IFN-inducible gene Mx was up-regulated in the gsIFN-overexpressing cells after GSIV infection. The virus load and titer were significantly reduced compared with that in control cells. Additionally, a lower level of virus major capsid protein synthesis was confirmed by immunofluorescence assay compared to the control cells. These results suggest that the gsIFN gene plays an important role in the antiviral innate immune response. Copyright © 2015 Elsevier Ltd. All rights reserved.
    Molecular Immunology 02/2015; 65(2):350-359. DOI:10.1016/j.molimm.2015.02.015 · 2.97 Impact Factor
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    ABSTRACT: Chinese giant salamander hemorrhage is a newly emerged infectious disease in China and has caused huge economic losses. The causative pathogen has been identified as the giant salamander iridovirus (GSIV). In this study, the immunological responses and protection in Chinese giant salamander immunized with β-propiolactone inactivated GSIV are reported. Red and white blood cell counting and classification, phagocytic activity, neutralizing antibody titration, immune-related gene expression and determination of the relative percent survival were evaluated after vaccination. The red and white blood cell counts showed that the numbers of erythrocytes and leukocytes in the peripheral blood of immunized Chinese giant salamanders increased significantly on days 4 and 7 post-injection (P < 0.01). Additionally, the differential leukocyte count of monocytes and neutrophils were significantly different compared to the control group (P < 0.01); the percentage of lymphocytes was 70.45 ± 7.52% at day 21. The phagocytic percentage and phagocytic index was 38.78 ± 4.33% and 3.75 ± 0.52, respectively, at day 4 post-immunization which were both significantly different compared to the control group (P < 0.01). The serum neutralizing antibody titer increased at day 14 post-immunization and reached the highest titer (341 ± 9.52) at day 21. The quantitative PCR analysis revealed that the immunization significantly up-regulated the expression of immune related genes TLR-9 and MyD88 the first two weeks after immunization. The challenge test conducted at day 30 post-injection demonstrated that the immunized group produced a relative survival of 72%. These results indicate that the inactivated GSIV could elicit significant non-specific and specific immunological responses in Chinese giant salamander that resulted in significant protection against GSIV induced disease.
    Veterinary Microbiology 11/2014; 174(3-4). DOI:10.1016/j.vetmic.2014.10.028 · 2.51 Impact Factor
  • Jie Ma · Lingbing Zeng · Yuding Fan · Yong Zhou · Nan Jiang · Qian Chen
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    ABSTRACT: The hemorrhagic disease of grass carp (Ctenopharyngodon idellus), caused by grass carp reovirus (GCRV), is the most severe disease of the fish that leads to huge economic losses. GCRV, belonging to the genus Aquareovirus of the family Reoviridae, has been classified into three genotypes based on their phylogenetic relationship. It is essential to develop an effective method to inhibit the replication of different genotypes of GCRV simultaneously. In this report, two multiple-shRNAs expression vectors, named pMultishVP2/2 and pMultishVP6/7, were generated and investigated. pMultishVP2/2 targeted the VP2 gene of GCRV-JX0901 (genotype I) and the VP2 gene of HGDRV (Hubei grass carp disease reovirus; genotype III). pMultishVP6/7 targeted the VP7 gene of GCRV-JX0901 and the VP6 gene of HGDRV. These two multiple-shRNAs expression vectors can simultaneously, significantly inhibit the replication of GCRV- JX0901 and HGDRV in vitro. Compared to the positive control, CPE induced by GCRV-JX0901 or HGDRV in cell transfected with shRNA transcribing vector was significantly delayed. The quantitative PCR analysis of the GCRV genomic RNA revealed that the pMultishVP2/2 could simultaneously inhibit the GCRV-JX0901 and HGDRV VP2 coding genes by 89.02% and 89.84%, respectively. The pMultishVP6/7 could simultaneously inhibit the GCRV-JX0901 VP7 coding gene and HGDRV VP6 coding gene by 80.63% and 86.78%, respectively. Furthermore, compared to the positive control, the indirect immunofluorescence assay and western blot demonstrated that the protein expression of the two genotypes of GCRV decreased significantly. The results in this study indicated that this multiple-shRNAs expression system could be used as a cross-reactive antiviral agent for treating the hemorrhagic disease of grass carp caused by multiple genotypes of GCRV.
    Virus Research 05/2014; 189. DOI:10.1016/j.virusres.2014.05.009 · 2.32 Impact Factor
  • Source
    Hui Zhang · Lingbing Zeng · Yuding Fan · Yong Zhou · Jin Xu · Jie Ma
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    ABSTRACT: A rapid and sensitive loop-mediated isothermal amplification (LAMP) assay for Cyprinid herpesvirus 2 (CyHV-2) detection in gibel carp was developed. Following cloning and sequencing of the putative DNA helicase gene of CyHV-2 isolate from China, a set of four specific primers was designed based on the sequence. The MgCl2 concentration and the reaction temperature were optimized to 6 mM, 64°C, respectively. LAMP products were detected by visual inspection of a color change due to addition of SYBR Green I stain. The specificity and sensitivity of the LAMP assay were determined. No cross-reaction was observed with other fish DNA viruses including eel herpesvirus, koi herpesvirus, and Chinese giant salamander iridovirus. The LAMP assay was found to be equally sensitive as nested PCR. A comparative evaluation of 10 fish samples using LAMP and nested PCR assays showed an overall correlation in positive and negative results for CyHV-2. These results indicate that the LAMP assay is simple, sensitive, and specific and has a great potential use for CyHV-2 detection in the laboratory and field.
    The Scientific World Journal 01/2014; 2014:716413. DOI:10.1155/2014/716413 · 1.73 Impact Factor
  • Yan Meng · Hui Zhang · Hongwei Liang · Lingbing Zeng · Hanbin Xiao · Congxin Xie
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    ABSTRACT: The Chinese giant salamander (Andrias davidianus) iridovirus (GSIV) is an emerging infectious pathogen responsible for severe hemorrhagic disease and high mortality in cultured Chinese giant salamanders. A loop-mediated isothermal amplification (LAMP) assay based on the major caspid protein (MCP) gene has been developed to detect this virus. Primer pairs for the LAMP assay were designed based on the GSIV MCP gene sequence. Amplification results indicate that under optimized conditions the LAMP assay has the ability to specifically detect the virus in both diseased animals and infected epithelioma papilloma cyprinid (EPC) cells. The assay was shown to be 10-fold more sensitive than nested PCR and was able to detect concentrations of 10(-9) (approximately 0.01pg/μL). The LAMP assay is relatively easy to perform in situ and the amplification products can be observed directly under UV light or via staining with SYBR Green I. The LAMP assay is also rapid and cost-effective. This study establishes the use of a LAMP assay for rapid detection of GSIV, which is a novel and important tool for the diagnosis of GSIV infection in laboratory or farmed Chinese giant salamanders.
    Journal of virological methods 09/2013; 194(1-2). DOI:10.1016/j.jviromet.2013.08.024 · 1.78 Impact Factor
  • Yuding Fan · Shujing Rao · Lingbing Zeng · Jie Ma · Yong Zhou · Jin Xu · Hui Zhang
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    ABSTRACT: A novel fish reovirus, Hubei grass carp disease reovirus (HGDRV, formerly grass carp reovirus strain 104, GCRV104), was isolated from diseased grass carp in China in 2009. Thereafter, the full genome sequence was determined. This reovirus was propagated in grass carp kidney cell line with typical cytopathic effect. The total size of the genome is 23,706 bp with a 51% G+C content and the eleven dsRNA segments encode twelve proteins (two proteins encoded by segment eleven). Nucleotide sequence similarity search using Blastn found no significant matches except for the segment two, which partially matched with RNA-dependent RNA polymerase (RdRp) from several viruses in the genera Aquareovirus and Orthoreovirus, of the Reoviridae family. At the amino acid level, seven segments (Seg-1 to Seg-6, and Seg-8) matched with species in Aquareovirus (with 15-46% identities) and Orthoreovirus (12-44%) genera, while for four segments (Seg-7, Seg-9, Seg-10, and Seg-11) no similarities in these genera were found. Conserved terminal sequences, 5'-GAAUU----UCAUC-3', were found in each HGDRV segment at the 5' and 3' ends, and the 5'-terminal nucleotides were different from any known species in Aquareovirus. Phylogenetic analysis based on RdRp amino acid sequences from Reoviridae showed that HGDRV clustered with aquareoviruses prior to joining a branch common with orthoreoviruses. Based on these observations, we propose that HGDRV is a new species of Aquareovirus that is distantly related to any known species within this genus.
    Journal of General Virology 07/2013; 94(Pt_10). DOI:10.1099/vir.0.054767-0 · 3.18 Impact Factor
  • Jin Xu · Lingbing Zeng · Hui Zhang · Yong Zhou · Jie Ma · Yuding Fan
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    ABSTRACT: An epizootic with severe mortality has emerged in cultured gibel carp, Carassius auratus gibelio, in China since 2009, and caused huge economic loss. The signs and epidemiology background of the disease were investigated. Parasite examination, bacteria and virus isolation were carried out for pathogen isolation. The causative pathogen was obtained and identified as Cyprinid herpesvirus 2 (CyHV-2) by experimental infection, electron microscopy, cell culture, PCR assay and sequence alignment, designated as CyHV-2-JSSY. Experimental infection proved the high virulence of CyHV-2-JSSY to healthy gibel carp. Electron microscopy revealed that the viral nucleocapsid was hexagonal in shape measuring 110-120nm in diameter with a 170-200nm envelope. The virus caused significant CPE in Koi-Fin cells at the early passages, but not beyond the fifth passages. Sequence alignment of the partial viral helicase gene (JX566884) showed that it shared 99-100% identity to the published sequences of other CyHV-2 isolates. This study represented the first isolation and identification of CyHV-2 in cultured gibel carp in China and laid a foundation for the further studies of the disease.
    Veterinary Microbiology 06/2013; 166(3). DOI:10.1016/j.vetmic.2013.05.025 · 2.51 Impact Factor
  • Jin Xu · Lingbing Zeng · Xiaosong Luo · Yao Wang · Yuding Fan · Shiyuan Gong
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    ABSTRACT: A reovirus, designated as CCRV-730, was isolated from channel catfish, Ictalurus punctatus, fingerlings suffering a severe hemorrhage in Hubei province in China. Experimental infection confirmed the pathogenicity of the virus and proved it to be the causative pathogen. Signs of the diseased channel catfish included abdominal distention, eyes bulging and hemorrhages of operculum, lower jaw, skin and fin bases. The CCK cell line was used for viral pathogen isolation. The electron microscopy observation of the virus infected cells revealed that there were a large number of reovirus-like particles measuring 60–70 nm in diameter in cytoplasm arrayed in crystalline. The viral genomic RNA was extracted and analyzed by SDS-PAGE and the complete S-class (S7–S11) segments of genome RNA were cloned and sequenced. The sequence alignment analysis of the S-class segments of CCRV-730 with corresponding sequences of other Aquareovirus members in the genetic sequence database of NCBI indicated that the virus had high similarity to grass carp reovirus, especially shared 99%–100% nucleotide sequence identity with the grass carp reovirus 873 strain (GCRV-873). The results implied that the genetic variation of GCRV-873 potentially arose in natural environment and resulted in the viral host conversion or expansion and made it pathogenic to channel catfish.
    Aquaculture 01/2013; s 372–375:39–44. DOI:10.1016/j.aquaculture.2012.10.011 · 1.88 Impact Factor
  • Jinfeng Zhang · Lingbing Zeng · Hui Zhang · Yong Zhou · Yi Xiao · Lan Su · Zhengyong Gao
    01/2013; 20(1):129-136. DOI:10.3724/SP.J.1118.2013.00129
  • Yuding Fan · Jin Xu · Xiaosong Luo · Yong Zhou · Yi Xiao · Lingbing Zeng
    01/2013; 18(1):38-47. DOI:10.3724/SP.J.1118.2011.00038
  • Yao Wang · Lingbing Zeng · Jin Xu · Yong Zhou · Yi Xiao
    01/2013; 37(1):117. DOI:10.3724/SP.J.1231.2013.38299
  • Zhengyong Gao · Lingbing Zeng · Yan Meng · Xiaoling Liu · Bo Zhang
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    ABSTRACT: To determine the pathogenic bacterium infecting giant salamander (Andrias davidianus). Bacterium was isolated from the liver of diseased Chinese giant salamander and identified by the Biolog Microbial Identification System and molecular biology method. Healthy Chinese giant salamander and crucian carp were used for experimental infection with bacterial suspension. A bacterial strain JZ01 was isolated and identified from diseased giant salamander. Infection with the bacterial suspension to healthy giant salamander could reproduce the diseased symptoms as occurred naturally and the same bacterium could be recovered from these infected giant salamanders. The isolated bacterium also has certain pathogenicity to crucian carp. Identification by the Biolog Microbial Identification System, and further 16S rDNA sequence and phylogenetic analysis demonstrated that the bacterium isolated from diseased giant salamander was Citrobacter freundii. The susceptibility test to antibiotics demonstrated that the bacterial strain JZ01 was susceptible to aztreonam, cefepime and cefotamine. Citrobacter freundii is a pathogen for cultured Chinese giant salamander.
    ACTA MICROBIOLOGICA SINICA 02/2012; 52(2):169-76.
  • Jie Ma · Weiming Wang · Lingbing Zeng · Yuding Fan · Jin Xu · Yong Zhou
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    ABSTRACT: Two DNA constructs targeting the grass carp reovirus (GCRV) RNA dependent RNA polymerase (RdRp) gene and outer capsid protein (OCP) gene that were each 64 bp in length were synthesized chemically and cloned into pSilencer2.1-U6 neo plasmid, named pSi-RdRp1286 and pSi-OCP117, respectively. After transfection of pSi-RdRp1286 and pSi-OCP117 plasmids into CIK cells, the inhibition of GCRV replication in the cells were detected by observing cytopathic effect (CPE), quantitating virus titers (TCID(50)/mL) and real-time quantitative RT-PCR analysis of viral RdRp and OCP genes. Five days after the cells were challenged with GCRV, both pSi-RdRp1286 and pSi-OCP117 reduced the viral titers by 5.47 lgTCID(50)/mL and 4.37 lgTCID(50)/mL, respectively. Compared to the positive control, CPE induced by GCRV in transfected cells was delayed and significantly less. Furthermore, the real-time quantitative RT-PCR analysis of the viral RdRp gene and OCP gene showed that the targeted gene expression were reduced by 89% and 73%, respectively. These results proved that the plasmid-transcribed shRNAs could inhibit effectively GCRV replication in CIK cells. These shRNAs provide potential tools for inhibiting GCRV infection and replication both in vitro and in vivo.
    Journal of virological methods 08/2011; 175(2):182-7. DOI:10.1016/j.jviromet.2011.05.008 · 1.78 Impact Factor
  • Yang Liu · Zemao Gu · Yunchao Zhang · Lingbing Zeng
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    ABSTRACT: Myxobolus shantungensis Hu, 1965 infects gill arches of common carp Cyprinus carpio haematopterus (Temminck and Schlegel), causing serious pathological effects on host fish. An inadequate original description and absence of molecular data make accurate early diagnosis challenging. To augment the original description, M. shantungensis is redescribed here using morphological and molecular biological methods. Mature spores of M. shantungensis were ellipsoidal or apple shaped in frontal view and lemon shaped in lateral view, averaging 8.2  ±  0.3 μm (8.0-9.0 μm) ×  10.1 ±  0.5 μm (9.2-11.1 μm)  ×  6.9  ±  0.3 μm (6.0-7.4 μm). Some spores had three to four "V"-shaped valve edge markings on the posterior of the spore. The two equal polar capsules were oval, measuring 4.3  ±  0.3 μm (3.8-5.0 μm)  ×  3.2  ±  0.2 μm (2.8-3.5 μm), situated at the anterior extremity of the spore. Polar filaments coiled with six to seven turns. Scanning electron microscopy revealed that the surface of mature spores of M. shantungensis was generally pitted with a number of irregular ridges in shape. M. shantungensis is also characterized on the molecular level using the sequence of the small subunit ribosomal RNA gene. A BLAST search revealed that this sequence did not match any available sequences in GenBank.
    Parasitology Research 05/2011; 109(6):1619-23. DOI:10.1007/s00436-011-2450-0 · 2.10 Impact Factor
  • Yuding Fan · Shuo Li · Jie Qi · Lingbing Zeng · Qiwang Zhong · Quanqi Zhang
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    ABSTRACT: The type II interleukin-1 receptor (IL-1RII) cDNA was cloned from Japanese flounder (Paralichthys olivaceus) by mRNA differential display (DD-PCR) and rapid amplification of cDNA ends (RACE). The full-length cDNA is 1793 bp in length, including a 100 bp 5'-terminal untranslated region (UTR), a 430 bp 3'-terminal UTR, and a 1263 bp open reading frame (ORF). The ORF encodes 420 amino acid residues with an estimated molecular mass of 46.65 kDa. The protein possesses signature features of the IL-R family, consisting of one N-terminal signal peptide, one transmembrane (TM) domain, two Ig-like domains in its extracellular region, one short cytoplasmic tail of 17 amino acids and four conserved proline residue sites. The predicted amino acid sequence has 65.3%, 52.5% and 51.6% identity with gilthead seabream, rainbow trout and Atlantic salmon IL-1RII, respectively. Phylogenetic analysis supported a close relation to mammalian IL-1RII. Reverse Transcription Polymerase Chain Reaction (RT-PCR) analysis indicated that the IL-1RII gene expression of Japanese flounder was weakly up-regulated and reached the peak expression in the kidney, spleen, and gill at 6 h after infection with Vibrio anguillarum, at 1.9, 2.0 and 1.5 times relative to the uninfected fish, respectively. These results suggest that IL-1RII has a signal transduction function and possibly plays a minor role in immune regulation against bacterial(s) infection in Japanese flounder.
    Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology 05/2010; 157(1):59-65. DOI:10.1016/j.cbpb.2010.05.001 · 1.55 Impact Factor

Publication Stats

42 Citations
34.09 Total Impact Points


  • 2010–2015
    • Chinese Academy of Fishery Sciences
      • Freshwater Fisheries Research Center
      北江, Zhejiang Sheng, China
  • 2011
    • Huazhong Agricultural University
      • College of Fisheries
      Wu-han-shih, Hubei, China