Isolierung und Charakterisierung von Iridoviren Tierärztliche Fakultät - Digitale Hochschulschriften der LMU - Teil 01/07
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We describe the isolation and characterization of an iridescent virus from commercially produced colonies of Gryllus bimaculatus in Germany, which showed apparent mortality. Transmission electron microscopy studies on adult cricket specimens revealed the paracrystalline assembly of icosahedral virus particles in the cytoplasm of hypertrophied abdominal fat body cells. The infecting agent could be cultivated in the lepidopteran cell line sf-9, where it caused cytopathogenic effects such as cell hypertrophy, cytoplasmic vacuolization, and cell death within 8 days postinfection. Negatively stained virus particles (n = 100) had dimensions of 172 +/- 6 nm (apex to apex) and 148 +/- 5nm (side to side). SDS-polyacrylamide gel electrophoresis of virus proteins showed more than 20 distinct polypeptides with a major species of approximately 50 kDa. Analysis of the restriction fragment length profiles from digestion of purified viral DNA with the endonucleases EcoRI, BamHI, and HindIII showed marked differences from the profiles of iridoviruses of lower vertebrates (genus Ranavirus), e.g., Rana esculenta Iridovirus and Frog virus 3. Restriction enzyme digests with the endonucleases MspI and HpaII indicated the lack of methylation of viral DNA. Polymerase chain reaction led to the amplification of a 420-bp gene fragment with 97 % sequence homology to the major capsid protein gene of Invertebrate iridescent virus 6 (IIV-6). The data indicate, that this new isolate, which is the first iridescent virus reported from Germany, belongs to the genus Iridovirus of the family Iridoviridae. It is proposed to be termed Gryllus bimaculatus iridescent virus (GbIV).
The investigation of four other iridescent-like viruses which were obtained in 1998, 1999 and 2000 from various tissues of three reptile species, e.g. Pogona vitticeps, Chamaeleo quadricornis, and Chamydosaurus kingii, revealed cytoplasmic desoxyriboviruses with the exact morphological and molecular biological characteristics of the formerly described GbIV. We conclude that all five isolates of iridescent-like viruses from Germany are identical and represent variants or strains of the type species of the genus Iridovirus, IIV-6. The data indicate, that GbIV, an invertebrate iridescent virus, is able to replicate in reptiles.
We describe the isolation and characterization of an iridescent virus from commercially produced colonies of Gryllus bimaculatus in Germany, which showed apparent mortality. Transmission electron microscopy studies on adult cricket specimens revealed the paracrystalline assembly of icosahedral virus particles in the cytoplasm of hypertrophied abdominal fat body cells. The infecting agent could be cultivated in the lepidopteran cell line sf-9, where it caused cytopathogenic effects such as cell hypertrophy, cytoplasmic vacuolization, and cell death within 8 days postinfection. Negatively stained virus particles (n = 100) had dimensions of 172 +/- 6 nm (apex to apex) and 148 +/- 5nm (side to side). SDS-polyacrylamide gel electrophoresis of virus proteins showed more than 20 distinct polypeptides with a major species of approximately 50 kDa. Analysis of the restriction fragment length profiles from digestion of purified viral DNA with the endonucleases EcoRI, BamHI, and HindIII showed marked differences from the profiles of iridoviruses of lower vertebrates (genus Ranavirus), e.g., Rana esculenta Iridovirus and Frog virus 3. Restriction enzyme digests with the endonucleases MspI and HpaII indicated the lack of methylation of viral DNA. Polymerase chain reaction led to the amplification of a 420-bp gene fragment with 97 % sequence homology to the major capsid protein gene of Invertebrate iridescent virus 6 (IIV-6). The data indicate, that this new isolate, which is the first iridescent virus reported from Germany, belongs to the genus Iridovirus of the family Iridoviridae. It is proposed to be termed Gryllus bimaculatus iridescent virus (GbIV).
The investigation of four other iridescent-like viruses which were obtained in 1998, 1999 and 2000 from various tissues of three reptile species, e.g. Pogona vitticeps, Chamaeleo quadricornis, and Chamydosaurus kingii, revealed cytoplasmic desoxyriboviruses with the exact morphological and molecular biological characteristics of the formerly described GbIV. We conclude that all five isolates of iridescent-like viruses from Germany are identical and represent variants or strains of the type species of the genus Iridovirus, IIV-6. The data indicate, that GbIV, an invertebrate iridescent virus, is able to replicate in reptiles.
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