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  • 1
    Electronic Resource
    Electronic Resource
    Propagation of yellow grouper nervous necrosis virus (YGNNV) in a new nodavirus-susceptible cell line from yellow grouper, Epinephelus awoara (Temminck & Schlegel), brain tissue (2001)
    Oxford UK : Blackwell Science Ltd
    Journal of fish diseases 24 (2001), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: A nodavirus was isolated from diseased yellow grouper, Epinephelus awoara, larvae cultured in southern Taiwan. The histopathology and RT–PCR results confirmed that it was a fish nodavirus; its coat protein gene sequence was similar to that of red spotted grouper nervous necrosis virus (RGNNV) and it is named yellow grouper nervous necrosis virus (YGNNV). A new nodavirus-susceptible cell line, grouper brain (GB) was established and characterized from the brain tissue of yellow grouper. The GB cells multiplied well in Leibovitz’s L-15 medium supplemented with 10% foetal bovine serum at temperatures between 24 and 32 °C, and have been subcultured more than 80 times, becoming a continuous cell line. The GB cell line consists of fibroblast-like cells and some epithelioid cells. The cell line yielded titres of YGNNV up to 108.5 TCID50 mL–1. The GB cells effectively replicated the virus at 28 °C, which could be purified to homogeneity by caesium chloride gradient centrifugation. Electron microscopy studies showed that purified virus particles were 25–30 nm in diameter. The cytoplasm of infected cells was filled with aggregates of virus particles. These results indicate that the GB cell line is a significant tool for the study of fish nodaviruses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2001.00303.x
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  • 2
    Electronic Resource
    Electronic Resource
    In vitro neutralization by monoclonal antibodies against yellow grouper nervous necrosis virus (YGNNV) and immunolocalization of virus infection in yellow grouper, Epinephelus awoara (Temminck & Schlegel) (2001)
    Oxford UK : Blackwell Science Ltd
    Journal of fish diseases 24 (2001), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mouse monoclonal antibodies (MAbs) were produced by using yellow grouper nervous necrosis virus (YGNNV) as an immunogen, isolated from infected yellow grouper, Epinephelus awoara (Temminck & Schlegel), and propagated in GB cells. In enzyme linked immunosorbent assay (ELISA), 43 hybridoma clones secreting MAbs strongly reacted with the purified virus. Ten of them showed a higher neutralization index (NI) value between 6.5 and 4.5 (log10 NI) than the other 33 MAbs against YGNNV infection in cell culture. All 10 MAbs belonged to the IgG isotype with a κ light chain and recognized the 42 kDa coat protein of YGNNV by Western blot analysis. Immunohistochemical results demonstrated that the viral signals co-located with pathological lesions observed in retina, brain and spinal cord. These results indicate that the MAbs are useful for confirmative diagnosis of YGNNV infection.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2001.00293.x
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  • 3
    Electronic Resource
    Electronic Resource
    Molecular characterization and pathogenicity of a grouper iridovirus (GIV) isolated from yellow grouper, Epinephelus awoara (Temminck & Schlegel) (2002)
    Oxford UK : Blackwell Science Ltd.
    Journal of fish diseases 25 (2002), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Molecular characterization was carried out on an iridovirus isolated from yellow grouper, Epinephelus awoara. The major capsid protein (MCP) gene was located, sequenced and compared with homologous genes from other iridoviruses. The nucleotide sequence is 1392 bases long and contains a single open reading frame beginning at an ATG codon from the 5′ end and terminating at a TAA codon at the 3′ end. The open reading frame encodes a protein of 463 amino acids with a predicted molecular weight of 50 272 Da. Pairwise amino acid alignments detected a high degree of sequence identity between grouper iridovirus (GIV) MCP and the homologous genes of other iridoviruses. The MCP gene of GIV was most similar to the MCP gene from frog virus 3 (FV3) with 70% nucleotide and 73% amino acid sequence identity. The predicted molecular weight of the protein of this gene is comparable with the apparent weight obtained by SDS–PAGE. Pathogenicity of the GIV was investigated in yellow grouper by intraperitoneal injection of 107 and 104 TCID50 virus. Cumulative mortalities reached 100% within 11 and 25 days post-infection, respectively, while no grouper died in the control group. The molecular studies demonstrated that GIV is a member of the genus Ranavirus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2002.00343.x
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  • 4
    Electronic Resource
    Electronic Resource
    Establishment of cell lines from a tropical grouper, Epinephelus awoara (Temminck & Schlegel), and their susceptibility to grouper irido- and nodaviruses (2003)
    Oxford, UK : Blackwell Science Ltd
    Journal of fish diseases 26 (2003), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Four tropical marine fish cell lines have been established from the eye, fin, heart and swim bladder of grouper, Epinephelus awoara (Temminck & Schlegel). Optimum media and temperature conditions for maximum growth were standardized. The eye and swim bladder cells were mostly epithelial, but the fin and heart cells were mostly fibroblastic. The viability of cells was 95% after 1 year of storage in liquid nitrogen (−196 °C). Besides these four cell lines, previously established grouper brain, kidney and liver cell lines were also used for a viral susceptibility study which showed that all the cell lines were sensitive to grouper iridovirus, whereas only brain, fin and liver cell lines were susceptible to the yellow grouper nervous necrosis virus (a nodavirus). Electron microscopy studies of the grouper irido- and nodaviruses in ultrathin sections of infected cells showed an abundance of viral particles in the cytoplasm of the virus-infected cells indicating the effective replication of these two viruses. It is suggested that these cell lines can be used for the isolation of putative fish specific viruses and provide a valuable tool to study the mechanisms of host–pathogen interactions. Furthermore, these cell lines upon transfection, using pEGFP-C1 and pEGFP-aMT2.5 (ayu metallothionein promoter), produced significant fluorescent signals indicating their utility for exogenous studies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2003.00434.x
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  • 5
    Electronic Resource
    Electronic Resource
    The production of a lymphokine (macrophage activating factor) by rainbow trout, Oncorhynchus mykiss (Walbaum), leucocytes stimulated with the extracellular products of Mycobacterium sp. (2001)
    Oxford UK : Blackwell Science Ltd
    Journal of fish diseases 24 (2001), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The production of macrophage activation factor (MAF) by rainbow trout, Oncorhynchus mykiss (Walbaum), head kidney leucocytes was examined after culturing in vitro with extracellular products (ECP) collected from Mycobacterium sp. Cultures of leucocytes were prepared from naive fish, or fish previously vaccinated with either the ECP or with formalin killed whole cell preparations (WC) of the bacterium. The cells were then incubated with the ECP in vitro and the ability of their supernatants to activate macrophages assessed. Macrophages from control fish were incubated with the supernatants, and their ability to reduce nitroblue tetrazolium (NBT) measured as an indicator of macrophage activation. Incubation of head kidney macrophages from naive fish directly with 1, 10 or 100 μg mL–1 of ECP for 48 h significantly enhanced macrophage activation compared with control macrophages. Vaccination of fish with either ECP or WC had no significant effect on the respiratory burst of control macrophages 4 weeks post-vaccination. By the eighth week, however, absorbance levels of respiratory burst reflecting both the primary (cells from vaccinated fish cultured in vitro with PBS) and the secondary (cells cultured in vitro with ECP) MAF responses of fish vaccinated with ECP and WC, had peaked and these were significantly different from the non-vaccinated controls. This activity had fallen to levels similar to control fish by week 12 for fish vaccinated with WC.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2001.00284.x
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  • 6
    Electronic Resource
    Electronic Resource
    Mass mortality associated with a Sphaerospora-like myxosporidean infestation in juvenile cobia, Rachycentron canadum (L.), marine cage cultured in Taiwan (2001)
    Oxford UK : Blackwell Science Ltd
    Journal of fish diseases 24 (2001), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Cultured cobia, Rachycentron canadum, of 45–80 g exhibited anaemia and ascites, and a mottled red and grey, extremely enlarged kidney with cream-coloured patches or spherical nodules. Cumulative mortality was about 90% within 1 month. Extrasporogonic or sporogonic stages of a myxosporean appeared in the blood, glomerulus, renal tubules and renal interstitium. The renal tubules were the main target tissue of the parasite and were completely occluded by sporogonic pseudoplasmodia at various degrees of maturity. Many sporogonic stages were attached to the brush border of the epithelium of the renal tubules. Mature spores were seen in the lumen of the tubules. They were elongated or spherical with numerous refractile granules in the cytoplasm. The polar filament formed 3–5 coils. No bacteria or viruses were isolated from the diseased fish. Based on the results of microbiological, histopathological and electron microscopical examinations, the cobia disease was believed to be caused by a Sphaerospora-like myxosporean. This is the first report of a myxosporean in cobia in aquaculture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2001.00287.x
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  • 7
    Electronic Resource
    Electronic Resource
    A Piscirickettsia salmonis-like organism in grouper, Epinephelus melanostigma, in Taiwan (2000)
    Oxford, UK : Blackwell Science Ltd
    Journal of fish diseases 23 (2000), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2000.00250.x
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  • 8
    Electronic Resource
    Electronic Resource
    Two iridovirus-susceptible cell lines established from kidney and liver of grouper, Epinephelus awoara (Temminck & Schlegel), and partial characterization of grouper iridovirus (2000)
    Oxford, UK : Blackwell Science Ltd
    Journal of fish diseases 23 (2000), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Two iridovirus-susceptible cell lines were established and characterized from grouper Epinephelus awoara kidney and liver tissues. These cell lines have been designated GK and GL, respectively. The cells multiplied well in Leibovitz's L-15 medium, supplemented with 10% foetal bovine serum, at temperatures between 20 and 32 °C, and have been subcultured more than 120 times, becoming continuous cell lines. The cell lines consist of a heterogeneous mixture of fibroblastic and epithelial cells. The viability of cells, stored frozen in liquid nitrogen (−196 °C), was 95% after 1 year. Chromosome morphologies of GK and GL cells were homogeneous. Both cell lines were susceptible to grouper iridovirus, and yielded high titres of up to 108 TCID50 mL−1. In addition, both cell lines effectively replicated the virus, which could be purified to homogeneity by cesium chloride gradient centrifugation. Electron microscopy studies showed that purified virus particles were 170±10 nm in diameter, and were hexagonal in shape. Virus-infected cells showed an abundance of virus particles inside the cytoplasm. These results show that the GK and GL cell lines effectively replicate grouper iridovirus, and can be used as a tool for studying fish iridoviruses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2000.00247.x
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  • 9
    Electronic Resource
    Electronic Resource
    Nocardiosis in sea bass, Lateolabrax japonicus, in Taiwan (2000)
    Oxford, UK : Blackwell Science Ltd
    Journal of fish diseases 23 (2000), S. 0 
    Details
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: An epizootic in pond cultured sea bass, Lateolabrax japonicus, was caused by Nocardia sp. in Taiwan, in September and October 1997. The cumulative mortality within 1 month was 17.5% (3500 out of 20 000 fish) and diseased fish were 7 months old with total lengths from 25 to 30 cm. Multiple, yellowish white nodules, 0.1–0.2 cm in diameter, were scattered in the gill, heart, liver, spleen and kidney. Histopathologically, typical granulomatous lesions appeared in those organs. The morphology of isolated bacteria from brain heart infusion (BHI) medium or Lowenstein–Jensen medium (LJM) were bead-like filaments, as shown by Ziehl-Neelsen's (ZN) staining method. The gross lesion and histopathological changes found in experimentally infected fish were similar to those in naturally infected fish. Based on the growth characteristics, morphological and biochemical properties of the bacterium, and histopathological changes, the isolated bacteria were identified as Nocardiaseriolae. This is the first report of N. seriolae-infected sea bass in aquaculture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2761.2000.00217.x
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  • 10
    Electronic Resource
    Electronic Resource
    Photoreflectance study of the surface state density and distribution function of InAlAs (2001)
    [S.l.] : American Institute of Physics (AIP)
    Journal of Applied Physics 89 (2001), S. 396-400 
    Details
    ISSN: 1089-7550
    Source: AIP Digital Archive
    Topics: Physics
    Notes: Photoreflectance is used to investigate the band gap, built-in electric field, and surface Fermi level of a series of lattice-matched In0.52Al0.48As surface-intrinsic n+ structures having different undoped layer thicknesses. Experimental results indicate that, although the built-in electric field depends on the undoped layer thickness, there is a range of thickness within which the surface Fermi level is weakly pinned. From the dependence of electric field and surface Fermi level on the undoped layer thickness, we can determine that the surface states distribute over two separate regions within the energy band gap. The densities of the surface states are evaluated as well. Moreover, the dependence of the built-in electric field on undoped layer thickness is converted into the dependence of surface state density on the surface Fermi level in order to theoretically and exactly calculate the energy spectrum of the surface state density using a Guassian distribution function. The center and width of the distribution near the conduction band are obtained from the fitting parameters. © 2001 American Institute of Physics.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1063/1.1324682
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