Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals

Zhanjiang, China

Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals

Zhanjiang, China
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Li Y.,Guangdong Ocean University | Cai S.-H.,Guangdong Ocean University | Cai S.-H.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals
Current Microbiology | Year: 2011

A set of six specific primers was designed by targeting intergenic spacer region (IGS) sequences. With Bst DNA polymerase, the products could be clearly amplified for 60 min at 62°C in a simple water bath. The sensitivity of the loop-mediated isothermal amplification (LAMP) for detecting Metarhizium anisopliae var. anisopliae was about 0.01 pg fungal DNA per reaction (equivalent to 27 conidia). LAMP products could be judged with agar gel or naked eye after addition of SYBR Green I. There were no cross reactions with other fungal isolates indicating high specificity of the LAMP. The LAMP could detect the presence of M. anisopliae var. anisopliae from soil. The detection limits for M. anisopliae var. anisopliae of LAMP reaction was 50 conidia per reaction in soil. © 2011 Springer Science+Business Media, LLC.


Yang S.-P.,Guangdong Ocean University | Yang S.-P.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals | Wu Z.-H.,Guangdong Ocean University | Wu Z.-H.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals | And 2 more authors.
Current Microbiology | Year: 2011

Populations of marine red yeast from shrimps and the environments of shrimp culture were investigated from various areas at Zhanjiang in China. All strains were studied for the production of biomass and carotenoids. We isolated 88 marine red yeast strains and the average populations of marine red yeast in seawater and the water from shrimp culture ponds were 70.0 and 172.4 CFU per 100 ml water, respectively. For shrimp samples, average populations of marine red yeast from gills, intestines, and stomachs were 178.0, 15.0, and 8.0 CFU per shrimp, respectively. The isolates were grouped into nine species belonging to three genera as follows: Rhodosporidium, Rhodotorula, and Sporidiobolus. R. sphaerocarpum had the highest average biomass yield (10.3 ± 0.88 g/l), followed by S. ruineniae (10.1 g/l) and Rh. mucilaginosa (9.9 ± 1.75 g/l). R. paludigenum had the highest average carotenoid yield (2.83 ± 0.589 mg/l), followed by S. pararoseus (2.72 mg/l) and R. sphaerocarpum (2.59 ± 0.454 mg/l). The results showed that marine red yeasts were normal microbial components in the environments of shrimp culture and shrimps, and carotenoids are abundant in these marine red yeast. © 2011 Springer Science+Business Media, LLC.


Li Y.,Guangdong Ocean University | Cai S.-H.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals
Current Microbiology | Year: 2011

Thirty-six strains, numbered from PY01 to PY36, were isolated from six moribund Oreochromis niloticus. The biochemical characteristics of all strains conformed to the species description of Aeromonas sobria on the basis of API 20E and Biolog GN system. Furthermore, gyrB sequence of strain PY36 was sequenced and showed high similarity (99.8%) with A. sobria in Genbank. Antibiotic-resistance of strain PY36 was assessed by the Kirby-Bauer disk diffusion method, and the results showed it was susceptible and moderately susceptible to 12 and 3 of the 19 antimicrobials tested. Virulence of strain PY36 to juvenile tilapia was also tested, and we found that LD 50 was about 4.17 × 10 3 CFU per fish in intraperitoneal injection. This is the first article to report that A. sobria was the pathogenic agent of tail-rot disease in juvenile tilapia. A. sobria was multi-resistant to the most frequently used antimicrobial drugs in China, so the antimicrobial resistance test should be carried out when these bacteria are isolated from biological samples in order to avoid therapeutic failures and spread of the pathogenic organisms in the environment. © 2010 Springer Science+Business Media, LLC.


Liang H.Y.,CAS South China Sea Institute of Oceanology | Liang H.Y.,University of Chinese Academy of Sciences | Liang H.Y.,Guangdong Ocean University | Liang H.Y.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals | And 11 more authors.
Letters in Applied Microbiology | Year: 2010

Aims: The main aims of this study were to clone and express flagellin flaA gene from Vibrio alginolyticus strain HY9901, also to prepare mouse anti-FlaA polyclonal antibody for future pathogen or vaccine study. Methods and Results: The full-length flaA gene was amplified by PCR with designed primers. The open reading frame of flaA gene contains 1131 bp, and its putative protein consists of 376 amino acid residues. Alignment analysis indicated that the FlaA protein was highly conserved. SDS-PAGE indicated that the FlaA protein was successfully expressed in Escherichia coli BL21 (DE3). Then, the recombinant FlaA protein was purified by affinity chromatography, and the mouse anti-FlaA serum was produced. The expression of flaA gene was verified by various immunological methods, including western blotting, enzyme-linked immunosorbent assay (ELISA) and immunogold electron microscopy (IEM). Conclusions: Flagellin flaA gene was cloned and identified from V. alginolyticus HY9901, the recombinant FlaA protein was expressed and purified, and high-titre FlaA protein-specific antibody was produced. Western blot analysis revealed that the prepared antiserum not only specifically react to FlaA fusion protein, but also to natural FlaA protein of V. alginolyticus. The expressed FlaA protein was demonstrated, for the first time, as the component of flagella from V. alginolyticus by IEM. Significance and Impact of the Study: This study may offer important insights into the pathogenesis of V. alginolyticus, provide a base for further studies on the diagnosis and evaluation that whether the FlaA protein could be used as an effective vaccine candidate against infection by V. alginolyticus and other Vibrio species. Additionally, the purified FlaA protein and polyclonal antibody can be used for further functional and structural studies. © 2009 The Society for Applied Microbiology.


Xia L.,Guangdong Ocean University | Xia L.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals | Tong B.,Guangdong Ocean University | Xu L.,Guangdong Ocean University | And 5 more authors.
Journal of Fisheries of China | Year: 2017

Nocardia seriolae is the main pathogen of fish nocardiosis, which is a chronic systemic granuloma disease of fish. A gene of protein tyrosine phosphatase (PTP) was found by analyzing the whole genome sequence of N. seriolae, and bioinformatics analysis showed that the PTP gene mayencode a secreted protein which could possibly target host cell mitochondria. In this study, the gene cloning, subcellular localization, over-expression and mitochondrial membrane potential detection were carried out. The results showed that the protein PTP was identified in the extracellular products of N. seriolae, which confirmed that PTP was a secreted protein. Subcellular localization of PTP-GFP fusion proteins were evenly distributed in the whole cell of FHM cells, and did not coincide with the distribution of mitochondria, which indicated that the protein PTP was not targeted at mitochondria. Typical apoptotic features, such as nuclear pyrosis and apoptotic bodies, were found when PTP protein was expressed in FHM cells by both subcellular localization and over-expression studies. The mitochondrial membrane potential was significantly damaged in FHM at 48h after the transfection of pcDNA-PTP, which confirmed that PTP was a bacterial protein which can likely induce cell apoptosis. The gene cloning and preliminary function study of PTP from N. seriolae have laid the foundation for further research on the gene and for promoting the understanding of the pathogenic mechanism of N. seriolae.


Cai S.-H.,Guangdong Ocean University | Cai S.-H.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals | Cai S.-H.,Key Laboratory of Control for Diseases of Aquatic Economic Animals of Guangdong Higher Education Institutes | Lu Y.-S.,Guangdong Ocean University | And 20 more authors.
Diseases of Aquatic Organisms | Year: 2013

The outer membrane proteins of Vibrio alginolyticus play an important role in the virulence of the bacterium and are potential candidates for vaccine development. In the present study, the ompW gene was cloned, expressed and purified. A DNA vaccine was constructed by inserting the ompW gene into a pcDNA plasmid. Crimson snapper Lutjanus erythropterus (Bloch) were injected intramuscularly with the recombinant plasmid pcDNA-ompW. The expression of the DNA vaccine was detected in gill, head kidney, heart, liver, spleen and injection site muscle of crimson snapper by RT-PCR 7 and 28 d post-vaccination. The ELISA results demonstrated that the DNA vaccine produced an observable antibody response in all sera of the vaccinated fish. In addition, crimson snapper immunized with the DNA vaccine showed a relative percentage survival (RPS) of 92.53%, indicating effective protection against V. alginolyticus infection. © 2013 Inter-Research.


Cao Y.T.,CAS South China Sea Institute of Oceanology | Cao Y.T.,Guangdong Ocean University | Cao Y.T.,Guangdong Prov Key Laboratory Of Pathogenic Biology And Epidemiology For Aquatic Economic Animals | Cao Y.T.,University of Chinese Academy of Sciences | And 6 more authors.
Letters in Applied Microbiology | Year: 2010

Aims: The purpose of this study was to establish a loop-mediated isothermal amplification (LAMP) method for the rapid, sensitive detection of Vibrio harveyi in mariculture shellfish. Methods and Results: A set of four primers, two outer and two inner primers, were designed from the toxR gene sequence of V. harveyi. The LAMP reaction was conducted at 65°C for 60 min. There were no cross-reactions with other bacterial strains indicating a high specificity of the LAMP. The detection sensitivity of the LAMP assay for V. harveyi with both of pure cultures and added shellfish cultures is about 10-5 dilution level (equivalent to 17·2 cells per reaction). The amplification products were detected by visual inspection using SYBR Green I. The detection sensitivity using the LAMP method was 10 times higher than that of conventional PCR. Conclusions: The LAMP assay established in this study is an extremely specific, sensitive and rapid for identification of V. harveyi in mariculture shellfish. Significance and Impact of the Study: This LAMP technique provides an important detecting tool for the detection of V. harveyi infection both in the laboratory and field. This technique is recommended as an applied protocol for health management programme and disease surveillance of in hatcheries as well as in grow-out pond, to prevent the disease outbreak. © 2010 The Society for Applied Microbiology.

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