Search

Viruses and high temperature (HT) are the primary threats to silkworms. Changes in the expression of stress-response genes can be measured using quantitative polymerase chain reaction (qPCR) after exposure to viruses or HT. However, appropriate reference genes (RGs) for qPCR data normalization have not been established in this organism. In this study, we summarized the RGs used in the previous silkworm studies after infection with Bombyx mori nucleopolyhedrovirus (BmNPV), B. mori cytoplasmic polyhedrosis virus (BmCPV), or B. mori densovirus (BmDNV) or after HT treatment. The expression levels of these RGs were extracted from silkworm transcriptome data to screen for candidate RGs that were unaffected by the experimental conditions. Actin-1 ( A1), actin-3 ( A3), glyceraldehyde-3-phosphate dehydrogenase ( GAPDH), and translation initiation factor 4a ( TIF- 4A) were selected for further qPCR verification. The results of RNA-seq and qPCR showed that GAPDH and TIF- 4A were suitable RGs after BmNPV challenge or HT stress, whereas TIF- 4A was an appropriate RG for BmCPV or BmDNV-Z challenge in silkworms. These results suggested that TIF-4A may be the most appropriate RG for gene expression analysis after challenge with viruses or HT in silkworms. [ABSTRACT FROM AUTHOR]

In this study, we isolated and characterized an insect nidovirus from the mosquito Culex tritaeniorhynchus Giles (Diptera: Culicidae) in Vietnam, as an additional member of the new family Mesoniviridae in the order Nidovirales. The virus, designated 'Dak Nong virus (DKNV),' shared many characteristics with Cavally virus and Nam Dinh virus, which have also been discovered recently in mosquitoes, and these viruses should be considered members of a single virus species, Alphamesonivirus 1. DKNV grew in cultured mosquito cells but could not replicate in the cultured vertebrate cells tested. N-terminal sequencing of the DKNV structural proteins revealed two posttranslational cleavage sites in the spike glycoprotein precursor. DKNV is assumed to be a new member of the species Alphamesonivirus 1, and the current study provides further understanding of viruses belonging to the new family Mesoniviridae. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the occurrence of baculovirus infections in mosquitoes and characterize them by using molecular tools. Fortnightly collections were made of mosquito larvae in the city of Caraguatatuba. Six larvae of Culex quinquefasciatus were isolated that had white cysts (nodules) in epithelia cells of the posterior midgut, indicative of infection by a baculovirus. These larvae were subjected to DNA extraction. DNA was amplified, producing a fragment of around 600 nt of the lef-8 gene and 400 nt of Pif-2 gene. The sequences were aligned, using ClustalX 2.0, with partial sequences of lef-8 genes of baculoviruses isolated from members of other insect orders taken from the GenBank database and edited, and phylogenetic analysis was performed. Phylogenetic analysis performed with the lef-8 and pif-2 genes demonstrated that the baculovirus identified in Culex quinquefasciatus in the Caraguatatuba region is most closely related to the deltabaculovirus Culex nigripalpus nucleopolyhedrovirus. [ABSTRACT FROM AUTHOR]

Summary. The biological function of entomopoxvirus (EPV) spindles (inclusion bodies that lack virions), has not been elucidated. We characterized the function of EPV spindles in the cupreous chafer, Anomala cuprea (Coleoptera: Scarabaeidae). Spheroids or spheroids mixed with spindles of Anomala cuprea EPV were administered per os to the A. cuprea larvae. A significant increase in infectivity was induced by the addition of spindles to the spheroids, strongly suggesting that the spindles play an important role in the infection of the host insect. [ABSTRACT FROM AUTHOR]

Summary. A total of 30 iridoviruses collected from Australia, South-East Asia, North America, South America and Europe were characterised. With the exception of the South-East Asian iridoviruses all viruses were found to belong to the genus Ranavirus. All viruses, except those originating from South-East Asia, cross-reacted with antisera against epizootic haematopoietic necrosis virus (EHNV). Viruses or virus-infected cells were examined using electron microscopy, SDS PAGE, restriction endonuclease (RE) digestion, DNA hybridisation, and DNA sequencing. Data from RE digestion of genomic DNA, and from the sequencing of PCR products indicated that the viruses generally grouped according to their geographic and taxonomic (i.e. amphibian or fish) origin. The one exception to this was the viruses from the United Kingdom that grouped with the North American ranaviruses. The differences between specified genomic regions were small. To assess the validity of the differences in sequence homology, similar studies were performed with different isolates from two viruses (EHNV and Guatopo virus (GV), collected from different animals at different locations and time). The sequence data showed complete homology for the isolates for any one virus over the 200 and 586 bp regions examined. Collectively, the data showed that the coding region for the major coat protein (MCP) is stable for any one species (e.g. EHNV). [ABSTRACT FROM AUTHOR]