rhabdovirus
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Rapid antigen detection enzyme-linked immunosorbent assay (ELISA) testing of cell cultures with organ homogenate from fish, collected from farms with a predominance of common carp or in natural aquaculture in the Czech Republic between 1995 and 2008, identified piscine vesiculovirus in 27 of 178 samples. Using reverse transcription semi-nested PCR, targeting a 550 nucleotide region of the glycoprotein (G) gene, piscine vesiculovirus was confirmed in 23 of the 27 organ samples diagnosed by ELISA as infected. PCR products were amplified and sequenced from 18 isolates from common carp Cyprinus carpio (family Cyprinidae), 2 isolates from northern pike Esox lucius (family Esocidae), and 1 isolate each from Siberian sturgeon Acipenser baerii (family Acipenseridae), common barbel Barbus barbus (family Cyprinidae), and koi carp Cyprinus carpio koi (family Cyprinidae). The sequences (based on 401 nucleotides) clustered into 2 genogroups. The majority of isolates (n = 22), including those from sturgeon and pike, grouped with the spring viraemia of carp virus (SVCV) Genogroup I and Subgroup Id. The 22 isolates could be further subdivided into 2 groups: Id1 (n = 20) and Id2 (n = 2). A marker (a non-conservative nucleotide substitution) for the Id1 SVCV group was identified. It was specifically found in all sequences of Id1 isolates when testing SVCV originating from different countries. The remaining isolate from barbel, was classified in the pike fry-like rhabdovirus Genogroup IV. This is the first confirmation of natural SVCV infection in sturgeon and pike, and pike fry-like rhabdovirus infection in barbel. In the case of the pike fry-like rhabdovirus, this is also its first identification in the Czech Republic. According to the presence/absence of evident clinical signs of rhabdoviral disease in the 3 infected hosts, only the sturgeon seemed to be susceptible to the monitored rhabdovirus.
- MeSH
- fylogeneze MeSH
- infekce viry z čeledi Rhabdoviridae veterinární virologie MeSH
- nemoci ryb virologie MeSH
- Rhabdoviridae genetika izolace a purifikace MeSH
- ryby MeSH
- sekvence nukleotidů MeSH
- Vesiculovirus MeSH
- vodní hospodářství MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A virome screen was performed on a new breeding line, KB1, of blackcurrant. Rhabdovirus-like particles were observed by electron microscopy in ultrathin sections of flower stalks, and the complete genome sequence of a novel virus, provisionally named blackcurrant rhabdovirus 2 (BCRV2), was determined and verified using high-throughput sequencing. The genomic organization of BCRV2 was characteristic of cytorhabdoviruses (family Rhabdoviridae) and included seven genes: 3 ́- N-P ́-P-P3-M-G-L -5 ́. BLASTP analysis revealed that the putative L protein had the highest amino acid sequence identity (75 %) with strawberry virus 2. BCRV2 was detected in Cryptomyzusgaleopsidis, but efficient transmission by this aphid was not confirmed. Of note, we observed coinfection of the KB1 line with blackcurrant-associated rhabdovirus (BCaRV) by RT-PCR. This is likely the first evidence of the presence of a cyto- and a nucleorhabdovirus in a single host.
Virus diseases of strawberry present several complex problems. More than 25 viruses have been described in the genus Fragaria thus far. Here, we describe a novel rhabdovirus, tentatively named strawberry virus 1 (StrV-1), that infects F.ananassa and F.vesca plants. Genomic sequences of three distinct StrV-1 genotypes co-infecting a single F.ananassa host were obtained using combined Illumina and Ion Proton high-throughput sequencing. StrV-1 was transmitted to herbaceous plants via Aphisfabae and A.ruborum, further mechanically transmitted to Nicotianaoccidentalis 37B and sub-inoculated to N.benthamiana, N.benthamiana DCL2/4i, N.occidentalis 37B, and Physalisfloridana plants. Irregular chlorotic sectors on leaf blades and the multiplication of calyx leaves seem to be the diagnostic symptoms for StrV-1 on indexed F.vesca clones. StrV-1 was detected in asymptomatic grafted plants and in 49 out of 159 field strawberry samples via RT-PCR followed by Sanger sequencing. The bacilliform shape of the virions, which have a cytoplasm-limited distribution, their size, and phylogenetic relationships support the assignment of StrV-1 to a distinct species of the genus Cytorhabdovirus. Acyrthosiphonmalvae, A.fabae, and A.ruborum were shown to transmit StrV-1 under experimental conditions.
- MeSH
- fylogeneze MeSH
- genom virový MeSH
- jahodník virologie MeSH
- listy rostlin virologie MeSH
- mšice fyziologie virologie MeSH
- nemoci rostlin parazitologie virologie MeSH
- Rhabdoviridae klasifikace genetika izolace a purifikace MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The genus Cytorhabdovirus includes plant viruses with an unsegmented, single-stranded, negative-sense RNA genome that infect various plant hosts. In this work, we report the detection of a new cytorhabdovirus infecting elderberry (Sambucus nigra L.). Total RNA was purified from infected leaves and, after ribodepletion, sequenced using an Illumina system. The RNA genome of viral isolate B15 is 12,622 nucleotides (nt) long, and that of isolate B42 is 12,621 nt long. A nearly complete sequence (12,592 nt) was also obtained for a third isolate (B160). The RNA genomes of all three isolates showed an organisation typical of cytorhabdoviruses, harbouring all six of the expected genes (3 ́ N-P-P3-M-G-L 5 ́), separated by intergenic regions. These isolates were closely related to each other (99.5-99.6% nt sequence identity) and showed the highest overall similarity to trichosanthes associated rhabdovirus 1 (63.5% identity) and Wuhan insect virus 5 (58% identity), and similar results were obtained when comparing individual coding sequences or proteins. Phylogenetic analysis confirmed that this elderberry virus, for which we propose the name "sambucus virus 1" belongs to the genus Cytorhabdovirus and fulfils the criteria to represent a novel species.
- MeSH
- bez černý * MeSH
- bez * genetika MeSH
- fylogeneze MeSH
- genom virový MeSH
- nemoci rostlin MeSH
- otevřené čtecí rámce MeSH
- Rhabdoviridae * MeSH
- RNA MeSH
- virové proteiny genetika MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Sandflies are known vectors of leishmaniasis. In the Old World, sandflies are also vectors of viruses while little is known about the capacity of New World insects to transmit viruses to humans. Here, we relate the identification of RNA sequences with homology to rhabdovirus nucleocapsids (NcPs) genes, initially in the Lutzomyia longipalpis LL5 cell lineage, named NcP1.1 and NcP2. The Rhabdoviridae family never retrotranscribes its RNA genome to DNA. The sequences here described were identified in cDNA and DNA from LL-5 cells and in adult insects indicating that they are transcribed endogenous viral elements (EVEs). The presence of NcP1.1 and NcP2 in the L. longipalpis genome was confirmed in silico. In addition to showing the genomic location of NcP1.1 and NcP2, we identified another rhabdoviral insertion named NcP1.2. Analysis of small RNA molecules derived from these sequences showed that NcP1.1 and NcP1.2 present a profile consistent with elements targeted by primary piRNAs, while NcP2 was restricted to the degradation profile. The presence of NcP1.1 and NcP2 was investigated in sandfly populations from South America and the Old World. These EVEs are shared by different sandfly populations in South America while none of the Old World species studied presented the insertions.
- MeSH
- DNA MeSH
- leishmanióza * MeSH
- lidé MeSH
- Psychodidae * MeSH
- Rhabdoviridae * MeSH
- RNA MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Brazílie MeSH
- Jižní Amerika MeSH
A rabbit polyclonal antibody (PAb) raised against European catfish virus (ECV; isolated from black bullhead Ameiurus melas in France) was produced and then evaluated using a panel of 9 ranavirus isolates collected from different lower vertebrate species originating from Australia, North and South America, Southeast Asia, and Europe. Using ranavirus-infected epithelioma papillosum cyprini (EPC) cell cultures, the specificity of the PAb was determined by Western blot, immunogold electron microscopy, and direct enzyme-linked immunosorbent assay (ELISA). Western blot analysis demonstrated that the PAb reacted strongly with a protein with a molecular weight corresponding to approximately 49 kDa. Immunogold electron microscopy provided direct evidence that the epitopes recognized by this PAb were located on the outer surface of virions. The PAb was used for the preparation of a peroxidase-labeled conjugate for the direct ELISA detection of ranaviruses in infected EPC cell cultures. The specificity of the conjugated PAb was tested using ranaviruses, some representative fish viruses of the genera Rhabdovirus and Birnavirus, and samples from various non-infected fish species. The PAb detected all tested ranaviruses except for 2 Santee-Cooper ranaviruses. The direct ELISA enabled the detection of ranavirus from a concentration of 10(3.5) to 10(3.8) TCID50 ml(-1) cell culture. The results of this study revealed that the rabbit PAb raised against ECV could be useful for the development of specific and standardized diagnostic assays for the detection of ranaviruses from freshwater fish and amphibians.
Cimex lectularius, known as the common bed bug, is a widespread hematophagous human ectoparasite and urban pest that is not known to be a vector of any human infectious disease agents. However, few studies in the era of molecular biology have profiled the microorganisms harbored by field populations of bed bugs. The objective of this study was to examine the viruses present in a large sampling of common bed bugs and related bat bugs (Cimex pipistrelle). RNA sequencing was undertaken on an international sampling of > 500 field-collected bugs, and multiple workflows were used to assemble contigs and query these against reference nucleotide databases to identify viral genomes. Shuangao bed bug virus 2, an uncharacterized rhabdovirus previously discovered in Cimex hemipterus from China, was found in several bed bug pools from the USA and Europe, as well as in C. pipistrelle, suggesting that this virus is common among bed bug populations. In addition, Shuangao bed bug virus 1 was detected in a bed bug pool from China, and sequences matching Enterobacteria phage P7 were found in all bed bug pools, indicating the ubiquitous presence of phage-derived elements in the genome of the bed bug or its enterobacterial symbiont. However, viral diversity was low in bed bugs in our study, as no other viral genomes were detected with significant coverage. These results provide evidence against frequent virus infection in bed bugs. Nonetheless, our investigation had several important limitations, and additional studies should be conducted to better understand the prevalence and composition of viruses in bed bugs. Most notably, our study largely focused on insects from urban areas in industrialized nations, thus likely missing infrequent virus infections and those that could occur in rural or tropical environments or developing nations.
Carp from breeding strains with different genetic background present diverse levels of resistance to viral pathogens. Carp strains of Asian origin, currently being treated as Cyprinus rubrofuscus L., especially Amur wild carp (AS), were proven to be more resistant to koi herpesvirus disease (KHVD; caused by cyprinid herpesvirus 3, CyHV-3) than strains originating from Europe and belonging to Cyprinus carpio L., like the Prerov scale carp (PS) or koi carp from a breed in the Czech Republic. We hypothesised that it can be associated with a higher magnitude of type I interferon (IFN) response as a first line of innate defence mechanisms against viral infections. To evaluate this hypothesis, four strains of common carp (AS, Rop, PS and koi) were challenged using two viral infection models: Rhabdovirus SVCV (spring viremia of carp virus) and alloherpesvirus CyHV-3. The infection with SVCV induced a low mortality rates and the most resistant was the Rop strain (no mortalities), whereas the PS strain was the most susceptible (survival rate of 78%). During CyHV-3 infection, Rop and AS strains performed better (survival rates of 78% and 53%, respectively) than PS and koi strains (survival rates of 35% and 10%, respectively). The evaluation of virus loads and virus replication showed significant differences between the carp strains, which correlated with the mortality rate. The evaluation of type I IFN responses showed that there were fundamental differences between the virus infection models. While responses to the SVCV were high, the CyHV-3 generally induced low responses. Furthermore, the results demonstrated that the magnitude of type I IFN responses did not correlate with a higher resistance in infected carp. In the case of a CyHV-3 infection, reduced type I IFN responses could be related to the potential ability of the virus to interfere with cellular sensing of foreign nucleic acids. Taken together, the results broaden our understanding of how common carp from different genetic strains interact with various viral pathogens.
- MeSH
- Herpesviridae fyziologie MeSH
- herpetické infekce imunologie veterinární MeSH
- infekce viry z čeledi Rhabdoviridae imunologie veterinární MeSH
- interferon typ I genetika imunologie MeSH
- kapři genetika imunologie MeSH
- nemoci ryb imunologie MeSH
- odolnost vůči nemocem genetika MeSH
- Rhabdoviridae fyziologie MeSH
- rybí proteiny genetika imunologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
