Guangdong Provincial Key Laboratory of Pathogenic Biology

Zhanjiang, China

Guangdong Provincial Key Laboratory of Pathogenic Biology

Zhanjiang, China

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Zhang X.Z.,CAS South China Sea Institute of Oceanology | Zhang X.Z.,University of Chinese Academy of Sciences | Zhang X.Z.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Zhang X.Z.,Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals | And 14 more authors.
Journal of Fish Diseases | Year: 2011

The heat-shock cognate 70 (HSC70) gene of humphead snapper, Lutjanus sanguineus, designated as ByHSC70, was cloned by rapid amplification of cDNA ends (RACE) with the primers designed from the known expressed sequence tag (EST) identified from the subtracted cDNA library of the head kidney of humphead snapper. The full-length cDNA of ByHSC70 is 2313bp, containing a 5' terminal untranslated region (UTR) of 96bp, a 3' terminal UTR of 267bp, and an open reading frame (ORF) of 1950bp encoding a polypeptide of 650 amino acids with a theoretical molecular weight of 71.21kDa and an estimated isoelectric point (pI) of 5.08. ByHSC70 contained three classical HSP70 family signatures. BLAST analysis showed that the amino acid sequence of ByHSC70 had the highest similarity of 99% when compared with other HSC70s. Fluorescent real-time quantitative RT-PCR was used to examine the expression of ByHSC70 gene in eight kinds of tissues/organs of humphead snapper after challenge with Vibrio harveyi. There was a clear time-dependent expression pattern of ByHSC70 in head kidney, spleen and thymus after bacterial challenge, and the expression of mRNA reached a maximum level at 9, 6 and 24h post-infection and then returned to control levels after 15, 24 and 36h, respectively. Our results suggest that HSC70 is an important component in the immune system of humphead snapper, its their rapid transcriptional upregulation in response to V. harveyi infection might be important for survival of humphead snapper. © 2011 Blackwell Publishing Ltd.


Zhou Z.,Guangdong Ocean University | Zhou Z.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Zhou Z.,Key Laboratory Of Control For Diseases Of Aquatic Economic Animals Of Guangdong Higher Edu Institute | Pang H.,Guangdong Ocean University | And 17 more authors.
Fish and Shellfish Immunology | Year: 2013

Type III secretion system (T3SS) in Vibrio alginolyticus is essential for its pathogenesis. VscO's homologous proteins FliJ, InvI and YscO have been suggested to be putative chaperone escorts although its function in V. alginolyticus is unclear. To investigate the physiological role of VscO, a mutant strain of V. alginolyticus with an in-frame deletion of the vscO gene was constructed in the present study. One finding was that the mRNA expression levels of SycD, VopB and VopD proteins decreased in the δ. vscO mutant. In addition, the δ. vscO mutant showed an attenuated swarming ability and a ten-fold decrease in the virulence to fish. However, the δ. vscO mutant showed no difference in the biofilm formation and ECPase activity. Complementation of the mutant strain with the vscO gene could restore the phenotypes of the wild-type strain. Finally, the recombinant VscO protein caused a high antibody titer and an effective protection against lethal challenge with the wild-type strain V. alginolyticus. These results indicated that VscO protein has a specific role in the pathogenesis of V. alginolyticus and it may be a candidate antigen for development of a subunit vaccine against vibriosis. © 2013 Elsevier Ltd.


Cai S.H.,Guangdong Ocean University | Cai S.H.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Cai S.H.,Economic Animals of Guangdong Higher Education Institutes | Lu Y.S.,Guangdong Ocean University | And 8 more authors.
Journal of Fish Diseases | Year: 2013

The outer membrane proteins of the marine aquatic animal pathogen, Vibrio alginolyticus, play an important role in the virulence of the bacterium and are potential candidates for vaccine development. In this study, the gene encoding an outer membrane protein-OmpU was cloned and expressed in Escherichia coli. Polyclonal antibodies were raised in rabbits against the purified recombinant OmpU, and the reaction of the antibody was confirmed by Western blotting using the isolated OmpU and the recombinant OmpU of V. alginolyticus. To analyze the immunogenicity of the recombinant OmpU, crimson snapper, Lutjanus erythropterus Bloch, were immunized by intraperitoneal injection, and antibody response was assessed by the enzyme-linked immunosorbent assay (ELISA). The results demonstrated that the recombinant OmpU produced an observable antibody response in all sera of the vaccinated fish. The vaccinated fish were challenged by virulent V. alginolyticus and observed to have high resistance to infection. These results indicate that the recombinant OmpU is an effective vaccine candidate against V. alginolyticus in L. erythropterus. © 2013 Blackwell Publishing Ltd.


Zhang X.,CAS South China Sea Institute of Oceanology | Zhang X.,University of Chinese Academy of Sciences | Zhang X.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Zhang X.,Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals | And 8 more authors.
Marine Genomics | Year: 2011

Heat shock protein 10 (HSP10) gene of humphead snapper (Lutjanus sanguineus), designated as ByHSP10, was cloned by rapid amplification of cDNA ends (RACE) techniques with the primers designed from the known EST sequence identified from the subtracted cDNA library of the head kidney of humphead snapper. Sequence analysis showed the full length cDNA of ByHSP10 was 529. bp, containing a 5' terminal untranslated region (UTR) of 51. bp, a 3' terminal UTR of 181. bp, and an open reading frame (ORF) of 297. bp encoding a polypeptide of 99 amino acids. Based on the deduced amino acid sequence, the theoretical molecular mass of ByHSP10 was calculated to be 10.92. kDa with an isoelectric point of 9.46. Moreover, chaperonins hsp10/cpn10 signature was found in the amino acids sequence of ByHSP10 by PredictProtein. BLAST analysis revealed that the amino acids of ByHSP10 had the highest homology of 88% compared with other HSP10s. Fluorescent real-time quantitative RT-PCR was used to examine the expression of ByHSP10 gene in eight kinds of tissues of humphead snapper after the challenge with Vibrio harveyi. There was a clear time-dependent expression pattern of ByHSP10 in head kidney, spleen and thymus after bacteria challenge. The expression of mRNA reached the maximum level at the time point of 9. h, 6. h and 24. h, respectively and then returned to control level in 36. h. The up-regulated mRNA expression of ByHSP10 in humphead snapper after bacteria challenge indicated that the HSP10 gene was inducible and might be involved in immune response. A phylogenetic tree was constructed based on the ORF nucleotide sequences of HSP10 for 30 species. The relatonships among them were generally in agreement with the traditional taxonomy which suggested that HSP10 genes could aid in the system classification research. © 2010 Elsevier B.V.


Yang S.-P.,Guangdong Ocean University | Yang S.-P.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Wu Z.-H.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Wu Z.-H.,Zhongkai University of Agriculture and Engineering | And 2 more authors.
Israeli Journal of Aquaculture - Bamidgeh | Year: 2013

The effects of orally-administered dry and live marine red yeast Rhodosporidium paludigenum on antioxidant-related gene expression in the hepatopancreas and hemocytes of Litopenaeus vannamei were investigated by RT-PCR. In the hepatopancreas of L. vannamei fed dry yeast, manganese superoxidate dismutase (SODMn) and catalase (CAT) were enhanced while glutathione peroxidase (GPx) and ferritin remained at the same levels as in the control group. In the hepatopancreas of L. vannamei fed live yeast, SODMn and ferritin were higher than in the control. In hemocytes of L. vannamei fed dry yeast, SODMn was lower, ferritin was similar, and CAT and GPx fluctuated, in comparison to the control. In hemocytes of L. vannamei fed live yeast, SODMn was lower, ferritin was higher, and CAT and GPx tended to be lower than in the control group. Results suggest that consumption of the marine red yeast, R. paludigenum, can stimulate antioxidant gene expression in the hepatopancreas of shrimp.


Liang H.,Guangdong Ocean University | Liang H.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Liang H.,Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals | Xia L.,Guangdong Ocean University | And 9 more authors.
Fish and Shellfish Immunology | Year: 2010

Vibrio alginolyticus is one of ubiquitous pathogens infecting human and marine animals. Flagellins of bacteria play an important role in infecting animals and inducing host immune response. In the present research, flagellin flaC gene of V. alginolyticus strain HY9901 was cloned and expressed. The open reading frame of flaC gene contains 1155 bp and the putative protein consists of 384 amino acid residues. Polyclonal antibodies were raised in mouse against the purified recombinant FlaC protein and the reaction of the antibody was confirmed by western blot analysis using the FlaC protein and crude protein extracts of V. alginolyticus. Red snapper (Lutjanus sanguineus) vaccinated with recombinant FlaC produced specific antibodies, and were highly resistant to infection by virulent V. alginolyticus. This study indicates that the conserved FlaC is an effective vaccine candidate against V. alginolyticus infection. © 2010 Elsevier Ltd.


Pang H.-Y.,Guangdong Ocean University | Pang H.-Y.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Pang H.-Y.,Economic Animals of Guangdong Higher Education Institutes | Zhang X.-Z.,Guangdong Ocean University | And 14 more authors.
Aquaculture Research | Year: 2013

Vibrio alginolyticus is one of the most serious diseases in cultured marine and freshwater fish and shellfish. The absence of suitable vaccine or virulent marker can be the bottleneck to control V. alginolyticus infection. In this study, immunoproteomic approaches were undertaken to study the immunogenicity of the whole-cell protein of V. alginolyticus HY9901. The whole-cell proteins were analysed by two-dimensional gel electrophoresis and subsequent immunoblotting using the rabbit anti-V. alginolyticus HY9901 serum. A total of 55 immunogenic proteins were identified by immunoproteomic analysis. Of the 55 proteins, 51 are specific immunoreactive proteins and four are nonspecific immunoreactive proteins. Furthermore, outer membrane protein N (spot 2) was used as immunogens to immunize Epinephelus coioides for investigation of protective abilities and activities. The E. coioides immunized with OmpN has abilities to fight against infections caused by V. alginolyticus. The other novel immunogenic proteins may be developed as alternative antigens for further study of V. alginolyticus vaccine and diagnostics. These data show that immunoproteomics methods can be successfully applied in identifying immunogenic proteins of V. alginolyticus, which helps to search for the protective antigens in future. © 2012 Blackwell Publishing Ltd.


Tang J.,Guangdong Ocean University | Tang J.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Tang J.,Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals | Cai J.,Guangdong Ocean University | And 17 more authors.
Fish and Shellfish Immunology | Year: 2014

The effects of a Chinese herbal mixture (CHM) composed of astragalus, angelica, hawthorn, Licorice root and honeysuckle on immune responses and disease resistant of Nile tilapia (Oreochromis niloticus GIFT strain) were investigated in present study. Fish were fed diets containing 0 (control), 0.5%, 1.0%, 1.5% or 2.0% CHM (w/w) for 4 weeks. And series of immune parameters including lysozyme, cytokine genes TNF-α and IL-1β, superoxide dismutase (SOD), peroxidase (POD), malondialdehyde (MDA) were measured during test period. After four weeks of feeding, fish were infected with Aeromonas hydrophila and mortalities were recorded. Results of this study showed that feeding Nile tilapia with CHM-supplementation diet stimulated lysozyme activity, SOD activity and POD activity in serum, induced TNF-α and IL-1β mRNA expression in head kidney and spleen, but decreased serum MDA content. All CHM-supplemental groups showed reduced mortalities following A.hydrophila infection compared with the group fed the control diet. These results suggested that this CHM can be applied as a tilapia feed supplement to elevate fish immunity and disease resistance against A.hydrophila. © 2014 Elsevier Ltd.


Gan Z.,Guangdong Ocean University | Gan Z.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Gan Z.,Key Laboratory of Control for Disease of Aquatic Animals of Guangdong Higher Education Institutes | Wang B.,Guangdong Ocean University | And 17 more authors.
Gene | Year: 2014

CD2BP2 (CD2 cytoplasmic tail binding protein 2), one of several proteins interacting with the cytoplasmic tail of CD2, plays a crucial role in CD2-triggered T cell activation and nuclear splicing. The studies on CD2BP2 have tended to be confined to a few mammals, and little information is available to date regarding fish CD2BP2. In this paper, a CD2BP2 gene (On-CD2BP2) was cloned from Nile tilapia, Oreochromis niloticus. Sequence analysis showed that the full length of On-CD2BP2 cDNA was 1429. bp, containing a 5'untranslated region (UTR) of 111. bp, a 3'-UTR of 193. bp and an open reading frame of 1125. bp which is encoding 374 amino acids. Two important structural features, a GYF domain and a consensus motif GPFXXXXMXXWXXXGYF were detected in the deduced amino acid sequence of On-CD2BP2, and the deduced genomic structure of On-CD2BP2 was similar to the known CD2BP2. The mRNA expression of On-CD2BP2 in various tissues of Nile tilapia was analyzed by fluorescent quantitative real-time PCR. In healthy Nile tilapia, the On-CD2BP2 transcripts were mainly detected in the head kidney and spleen. While vaccinated with inactivated Streptococcus agalactiae, the On-CD2BP2 mRNA expression was significantly up-regulated in the head kidney, spleen and brain 48. h post immunization. Moreover, there was a clear time-dependent expression pattern of On-CD2BP2 after immunization and the expression reached the highest level at 24. h in the brain and 48. h in the head kidney and spleen. This is the first report of proving the presence of a CD2BP2 ortholog in fish, and investigating its tissue distribution and expression profile in response to bacterial stimulus. These findings indicated that On-CD2BP2 may play an important role in the immune response to bacteria in Nile tilapia. © 2014 Elsevier B.V.


Cai J.,Guangdong Ocean University | Cai J.,Guangdong Provincial Key Laboratory of Pathogenic Biology | Cai J.,Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals | Wei S.,Guangdong Ocean University | And 20 more authors.
Marine Genomics | Year: 2013

It is well known that nonspecific cytotoxic cells (NCCs) are kinds of natural killer cell mediated innate immune responses in teleosts. The nonspecific cytotoxic cell receptor protein 1 (NCCRP-1) is an important cell surface protein on NCC, which serves crucial functions in target cell recognition and cytotoxicity activation. In the present study, a nonspecific cytotoxic cell receptor protein NCCRP-1 (Ls-NCCRP1) was cloned from red snapper, Lutjanus sanguineus. The Ls-NCCRP1 cDNA is composed of 986. bp with a 43. bp of 5'-UTR, 702. bp open reading frame (ORF) and 241. bp 3'-UTR, encoding a polypeptide of 233 amino acids (GenBank accession no: ADK32635). Phylogenetic analysis revealed that Ls-NCCRP1 showed highest similarity to sea bream NCCRP-1. Quantitative real-time PCR (qRT-PCR) analysis showed that Ls-NCCRP1 had relatively high expression level in the head kidney, spleen and liver. After Vibrio alginolyticus infection, transcripts of Ls-NCCRP1 increased and reached its peak at 4. h p.i. These results indicated that Ls-NCCRP1 may play an important role in innate immune response to bacteria. © 2013 .

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