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At least 19 recordsLinked to original sources

Broad geographical circulation of a novel vesiculovirus in bats in the Mediterranean region.

Bats are the natural reservoirs for a variety of emerging and re-emerging viruses. Among them, rabies virus (genus Lyssavirus, family Rhabdoviridae) is one of the first and most emblematic described in these animals. Since its first description, several new bat lyssaviruses have been regularly identified. In addition to lyssaviruses, other bat rhabdoviruses have also been discovered, including members of the genera Vesiculovirus, Ledantevirus and, more recently, Alphanemrhavirus and Tupavirus. However, the family Rhabdoviridae is one of the most abundant and diverse viral families, with 434 officially recognized species, divided into 5 subfamilies and 56 different genera. The number of rhabdoviruses associated with bats is therefore probably higher than that currently available. In this study, we first developed and validated a combined nested RT-qPCR technique (pan-rhabdo RT-nqPCR) dedicated to the broad detection of animal rhabdoviruses. After validation, this technique was used for a large retrospective screening of archival bat samples (n = 1962), including blood (n = 816), brain (n = 723) and oral swab (n = 423). These samples were collected from various bat species over a 12-year period (2007-2019) in 9 different countries in Europe and Africa. A total of 23 samples (1.2%) from bat species Miniopterus schreibersii, Rhinolophus euryale and Rhinolophus ferrumequinum tested positive for rhabdovirus infection, including 17 (2.1%) blood and 6 (1.4%) oral swab samples, all collected from bats originating from the Mediterranean region. Complete virus genome sequences were obtained by next-generation sequencing for most of the positive samples. Molecular and phylogenetic analysis of these sequences demonstrated that the virus isolates, named Mediterranean bat virus (MBV), were closely related and represented a new species, Mediterranean vesiculovirus, within the genus Vesiculovirus. MBV was more specifically related to other bat vesiculoviruses previously described from China and North America, together clustering into a distinct group of bat viruses within this genus. Interestingly, our results suggest that MBV is widespread, at least in the western part of the Mediterranean region, where it circulates in the blood of several bat species. These results expand the host range and viral diversity of bat vesiculoviruses, and pave the way for further studies to determine the transmission route and dissemination dynamics of these viruses in bat colonies, as well as to assess their potential threat to public health.

Animals↗

Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction.

We describe a reverse transcription-polymerase chain reaction (RT-PCR) and a nested-PCR for diagnosis of Piry, Carajás, Cocal, and Alagoas vesiculoviruses from Brazil. The RNA extracts of viral and clinical samples were submitted to a RT-PCR using Vesiculovirus G primers that amplify part of the glycoprotein gene. The RT-PCR produced amplicons of expected size, 290 base pair, for the four studied viruses. The RT-PCR showed a high sensitivity being 151.3 times (2.18 log) more sensitive for the detection of Piry virus than the classical procedure for virus detection in tissue culture based on the viral cytophatic effect. Amplicons had nucleotides sequenced and were aligned in order to select internal primers for a nested-PCR to confirm the origin of Piry, Carajás, Cocal, and Alagoas Vesiculovirus. Ten blood and tarsal pad epithelial samples of infected Guinea-pigs had Vesiculovirus genome amplified by RT-nested-PCR.

Animals↗

Genome RNA terminus conservation and diversity among vesiculoviruses.

Sequence analysis of the RNA genome termini of various vesiculovirus standard and defective interfering (DI) particles demonstrated that some virus regulatory sequences and domains of virus N protein are highly conserved while others show considerable divergence. Clearly, distinct RNA signal sequences and protein-coding regions of these virus genomes have quite different evolutionary pressures or constraints. Terminal regions of DI-particle RNA genomes of these viruses were found to possess self-complementary stems at the RNA termini, demonstrating the conservation of this DI-particle structural feature throughout the vesiculovirus group. A high degree of conservation of the 3'-terminal sequences of recent and historic isolates of vesicular stomatitis virus New Jersey was also demonstrated.

Amino Acid Sequence↗

Susceptibility of various animals to the vesiculovirus Piry.

To determine the pathogenic potential of the vesiculovirus Piry for domestic animals, two ponies, two steers, three sheep, three goats and three pigs were inoculated intradermally in the tongue or, in the case of the pigs, in the snout, heel and coronary band. Inoculated animals were housed in one room and allowed to mingle freely with an equal number of uninoculated contact animals of each species. Clinical signs of infection, consisting of elevated temperature and ulcers at the inoculation sites, were only observed in the ponies, but all inoculated animals developed specific antibody following inoculation. In addition, one of the contact sheep had neutralizing antibody to Piry at 7 and 29 days post inoculation suggesting a contact infection. Virus was not demonstrated in tissues, other than tongue, of any animal. The failure of Piry virus to produce lesions in steers, sheep, goats and pigs and only limited ulcerations in ponies suggests that this virus is not similar pathogenically to New Jersey and Indiana strains of vesiculoviruses which produce classical vesicular stomatitis. Lethal infections were produced by inoculation into suckling mice and hamsters, adult hamsters and embryonating chicken eggs. Further, lethal infections followed contact of adult female hamsters with their inoculated litters.

Animals↗

A newly recognized vesiculovirus, Calchaqui virus, and subtypes of Melao and Maguari viruses from Argentina, with serologic evidence for infections of humans and horses.

In 1983, 17 virus strains were isolated from mosquitoes collected during an outbreak of western equine encephalitis in Santa Fe Province, Argentina. Strains of western equine encephalitis, Venezuelan equine encephalitis, St. Louis encephalitis, and Antequera viruses were isolated, as were several bunyaviruses of the California and Bunyamwera serogroups and a new vesiculovirus. Complement fixation and neutralization tests were used to identify the California serogroup virus as a subtype of Melao virus, the Bunyamwera serogroup virus as a subtype of both Maguari and Playas viruses, and the vesiculovirus as a newly recognized agent for which the name Calchaqui virus is proposed. A limited serosurvey of horses and humans in Santa Fe Province and horses from the adjacent Santiago del Estero Province was performed to determine the prevalence of neutralizing antibody to the subtypes of Melao and Maguari viruses and to Calchaqui virus. The high prevalence of antibodies to these three agents indicates the need for further studies of their disease potential in horses, because they are closely related to several other viruses that are known equine pathogens.

Animals↗

Malpais spring virus: a new vesiculovirus from mosquitoes collected in New Mexico and evidence of infected indigenous and exotic ungulates.

Two virus isolates, 1 each from Aedes campestris and Psorophora signipennis mosquitoes collected in south central New Mexico in August 1985, were shown by neutralization tests to be identical to each other, but not to any of more than 250 arthropod-borne and other viruses. Electron microscopy of 1 isolate (85-488NM, chosen as the prototype) indicated that this strain shares morphologic characteristics with viruses of the family Rhabdoviridae. Indirect fluorescent antibody tests indicated that this virus is a member of the genus Vesiculovirus, but is not closely related to any of the North American or other rhabdoviruses with which it was tested, including vesicular stomatitis (Indiana) and vesicular stomatitis (New Jersey) viruses. The name Malpais Spring virus is proposed for this newly recognized vesiculovirus. A serologic survey indicated that Malpais Spring virus infects indigenous (mule deer and pronghorn) and exotic (gemsbok) ungulates at and near the sites where the mosquitoes from which the virus strains were isolated were collected. Antibody prevalence in wild animals indicates that the pronghorn and gemsbok may play roles as hosts for Malpais Spring, epizootic hemorrhagic disease (New Jersey), and bluetongue viruses in this area.

Aedes↗

Phlebotomine sand flies as vectors of vesiculoviruses: a review.

Phlebotomine sand flies appear to be involved in the epizootiology of 10 of the 16 currently recognized vesiculoviruses. Evidence in support of this includes isolation of viruses from wild-caught male and female sand flies; demonstration of oral infection, replication, and bite transmission of viruses by sand flies; the temporal and/or spatial association between infected sand flies and infected vertebrates; and demonstration of transovarial virus transmission. The present review summarizes the known sand fly-vesiculovirus relationships.

Animals↗

Complete genome sequences of Chandipura and Isfahan vesiculoviruses.

Chandipura virus (CHPV) and Isfahan virus (ISFV) are two members of the genus Vesiculovirus from Asia. Both are arthropod-transmitted and are able to infect humans, but neither causes vesicular stomatitis in livestock. The complete genome sequence for each virus has been determined. The negative-sense RNA genome comprises 11,119 nt (CHPV) or 11,088 nt (ISFV). The most variable of the non-transcribed regions is the intergenic spacer at the G-L gene junction (4 bases in ISFV, 20 in CHPV). Phylogenetic analysis of deduced protein sequences shows that although CHPV and ISFV are distinct viruses, they are more related to each other than either is to the New World vesicular stomatitis viruses (VSV). The South American virus, Piry virus, is more closely related to the Asian viruses ISFV and CHPV, than it is to VSV.

Amino Acid Sequence↗

Sequences of Chandipura virus N and NS genes: evidence for high mutability of the NS gene within vesiculoviruses.

The nucleotide sequence of the 3' end of the genome of Chandipura (CHP) virus, including the complete sequences of the nucleocapsid (N) and phosphoprotein (NS) genes was determined, principally from cloned cDNAs of the N and NS mRNAs. The NS mRNA of CHP virus is 908 bases in length and encodes a protein of 293 amino acids. Comparison of the CHP virus NS protein sequence with those of vesicular stomatitis virus of the New Jersey serotype (VSV (NJ)) and of the Indiana serotype (VSV (IND] revealed homologies of only 23 and 21%, respectively, with no consecutive stretches of more than four amino acids identical among the three sequences. As with the two VSV serotypes, the highest homology between the NS proteins of CHP and VSV was in a 20-amino acid region near the carboxy termini of the proteins. Of the potential phosphorylation sites, there are eight conserved serine or threonine residues among the three sequences. Despite the dissimilarity among primary sequences of the NS proteins, their overall structure, as assessed by amino acid composition and by the relative hydropathicities of the sequences, has been conserved throughout evolution. The N mRNA of CHP virus is 1291 bases long and encodes a protein of 422 amino acids. In contrast to the NS protein, the CHP N protein is at least 50% homologous to the N proteins of each of the VSV serotypes. We have identified a region near the center of these N protein sequences which is conserved among members of both the rhabdovirus and paramyxovirus families. This extent of conservation of the N protein sequences underscores the high rate of mutability of the NS protein sequences among the vesiculoviruses.

Amino Acid Sequence↗

Effect of melittin on transcription by vesiculovirus mutant and wild-type viruses.

The bee venom peptide melittin activated the virion transcriptase activity of three vesiculoviruses with preservation of virion structure. The kinetics of RNA synthesis were similar to those observed with purified transcribing nucleoprotein (TNP) preparations. Six temperature-sensitive host range (tdCE) mutants of Chandipura virus displayed 1.7- to 5.5-fold greater efficiencies of transcription at 39 degrees with melittin-permeabilized virions in comparison with TNP preparations. Comparative study of other host range mutants (tdCE3) and tsB1) of vesicular stomatitis virus (VSV) New Jersey and a thermosensitive polymerase mutant (tsG114) of VSV Indiana suggested that the enhanced transcription at 39 degrees associated with melittin-activated tdCE mutants was due to the retention of host factors in the virions.

Bee Venoms↗

The relationship of Piry virus to other vesiculoviruses: a re-evaluation based on the glycoprotein gene sequence.

By sequencing the 3 half of the Piry virus genome, we show that Piry virus, like the other vesiculoviruses, contains the genes for nucleoprotein N, phosphoprotein P, matrix protein M, glycoprotein G and polymerase protein L, in that order. Our analysis of the Piry G protein sequence suggests that Piry and Chandipura are related to each other as closely as the Indiana and New Jersey vesicular stomatitis virus serotypes are to each other. A re-examination of amino acid sequences in the nucleocapsid protein shows that this relationship is also true of the more conserved central region of this protein and that the greatest divergence between Piry and Chandipura has occurred in two other regions of the nucleocapsid protein.

Amino Acid Sequence↗

[Comparison of two neutralization tests, used in vesiculovirus Piry serodiagnosis].

A serological survey of Piry was undertaken of the sera of inhabitants of Catolância-Bahia State, Brazil. Serum results obtained by a vesiculovirus neutralization test of C6/36 cells read by ELISA are compared with those obtained by the classic technique, carried out on newborn mice. The agreement between the results was as high as 98.7% of the 204 sera tested and the neutralization test of C6/36 cells was chosen as the most suitable technique for the sero-survey testing.

Animals↗

Susceptibility of various animals to the vesiculoviruses Isfahan and Chandipura.

To determine the pathogenic potential of the vesiculoviruses Isfahan and Chandipura for domestic animals, two ponies, two steers, three sheep, three goats and three pigs were inoculated with each virus intradermally in the tongue or, in the case of the pigs, in the snout, heel and coronary band. The ponies were also inoculated intradermally in the right commissure of the mouth. Animals inoculated with each virus were housed in one room and allowed to mingle freely with an equal number of uninoculated contact animals of each species. Clinical signs of infection, consisting of ulcers at the inoculation sites, were observed in the Chandipura study in two inoculated ponies, one inoculated steer and one inoculated goat. No elevated temperature was observed. Virus was isolated from the ulcerated tongue tissue, but not from serial blood samples, oesophageal-pharyngeal mucus samples, or from the tissues which were collected at necropsy. Precipitating antibody was not detected by the immunoelectroosmophoresis (IEOP) test in any of the pre- or post-serum samples except from two inoculated sheep at 29 days post-inoculation (D.P.I.). Low levels of neutralizing activity were detected in pre-inoculation serum from all steers, pigs, contact sheep, and one contact goat. By 15 D.P.I. all inoculated animals and contact ponies and steers exhibited increased neutralizing antibody titres. In studies with the Isfahan virus, lesions developed only at the inoculation sites in the two ponies, and the virus was isolated. No virus was isolated from any blood, oesophageal-pharyngeal mucus samples or tissues collected at necropsy. All pre-inoculation sera were negative for neutralizing and precipitating antibodies. By 14 D.P.I. all inoculated animals exhibited neutralizing antibody, while all the contacts remained negative. The IEOP test remained negative for all animals throughout the experiment. A sub-passage of a suspension of Isfahan-infected tongue tissue injected into ponies and steers also yielded only firm swellings of lesser extent than the original reaction at the inoculation sites. With both viruses, lethal infections were produced by intracranial or intraperitoneal inoculation of day-old mice and hamsters, and by allantoic inoculation of embryonating chicken eggs. Adult mice, hamsters, guinea-pigs and rabbits produced serum antibodies but lacked clinical signs.

Animals↗

The sequence of the nucleocapsid protein (N) gene of Piry virus: possible domains in the N protein of vesiculoviruses.

A number of independent cDNA clones of the Piry virus N gene message were identified and sequenced. From the resulting sequences and previously published data, we derived the sequence of the mRNA for this protein. Sequence similarities of the translated region of Piry virus with that of other viruses suggest that Piry virus is as distantly related to Chandipura virus as it is to the vesicular stomatitis viruses of Indiana and New Jersey serotypes. Based on the relative conservation of the amino acid sequence of the nucleocapsid protein of these vesiculoviruses, the N protein can be subdivided into at least three regions, possibly indicative of functional domains.

Amino Acid Sequence↗

Linear mapping of tryptophan residues in Vesiculovirus M and N proteins by partial chemical cleavage.

Nonlimit chemical cleavage at tryptophan residues of protein labeled at the amino terminus afforded a simple procedure for generating specific fragments and for mapping tryptophan positions. A comparison of the matrix (M) and nucleocapsid (N) proteins of four members of the Vesiculovirus group by this procedure suggests considerable conservation of tryptophan number and location in the four serotypes examined.

Capsid↗

Carajas and Maraba viruses, two new vesiculoviruses isolated from phlebotomine sand flies in Brazil.

Two new rhabdoviruses, designated Carajas and Maraba, are described. Both were isolated from phlebotomine sand flies (Lutzomyia spp.) collected in the Amazon basin of Brazil. One recovery of Carajas virus was made from male sand flies. By complement-fixation and neutralization tests both agents were shown to be members of the vesicular stomatitis virus (VSV) serogroup (genus Vesiculovirus). The pathogenicity of the two viruses in mice and Vero cells is similar to that of VSV-Indiana and VSV-New Jersey. Both Carajas and Maraba viruses replicated in Lutzomyia longipalpis following intrathoracic inoculation, and both agents were transovarially transmitted in this sand fly species.

Animals↗