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1.
Artigo em Inglês | MEDLINE | ID: mdl-34463150

RESUMO

The circulation of arboviruses in livestock ruminants has often gone unrecognized owing to the fact that a significant percentage of arboviruses probably induce subclinical infections and/or negligible symptoms in infected animals. To determine the current situation of arbovirus circulation in the Yaeyama Islands, attempts to isolate viruses from bovine blood samples collected between 2014 and 2019 have been made. In total, 308 blood samples were collected during the study period, and 43 of them induced cytopathic effects (CPEs) in cell cultures. The identification of the CPE agents was performed by reported RT-PCR assays and a high-throughput analysis with a next-generation sequencing platform. The obtained viruses consisted of an orthobunyavirus (Peaton virus), Culicoides-borne orbiviruses (bluetongue virus serotypes 12 and 16, epizootic hemorrhagic disease virus [EHDV] serotypes 5, 6, and 7, D'Aguilar virus, and Bunyip Creek virus), and potential mosquito-borne orbiviruses (Yunnan orbivirus, Guangxi orbivirus, and Yonaguni orbivirus). Most of the orbiviruses were recovered from washed blood cells with mosquito cell cultures, suggesting that this combination was more efficient than other combinations such as plasma/blood cells and hamster cell lines. This marked the first time that the isolation of EHDV serotypes 5 and 6 and three potential mosquito-borne orbiviruses was recorded in Japan, showing a greater variety of orbiviruses on the islands than previously known. Genetic analysis of the isolated orbiviruses suggested that the Yaeyama Islands and its neighboring regions were epidemiologically related. Some of the viruses, especially the potential mosquito-borne orbiviruses, were isolated during several consecutive years, indicating their establishment on the islands.

2.
J Vet Med Sci ; 83(9): 1378-1388, 2021 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-34248104

RESUMO

Epizootic hemorrhagic disease (EHD) is an arthropod-borne disease of wild and domestic ruminants caused by the EHD virus (EHDV). To date, seven EHDV serotypes have been identified. In Japan, strain Ibaraki of EHDV serotype 2 has caused outbreaks of Ibaraki disease in cattle. In addition, EHDV serotype 7 (EHDV-7) has caused large-scale EHD epizootics. In mid-September 2016, eight cattle at a breeding farm in Fukuoka Prefecture, Japan developed fever. Since EHDV-7 was detected in sentinel cattle in western Japan in 2016, we suspected that the cause of this fever might be an EHDV-7 infection. In this study, we tested cattle for EHDV-7 and some other viruses. Consequently, EHDV was isolated from washed blood cells collected from three of the eight cattle, and genetic analysis of genome segment 2 revealed that this isolate was EHDV-7. Moreover, all affected cattle tested positive for anti-EHDV-7 neutralizing antibodies. Our results suggest that the fever was caused by EHDV-7 infection. In addition, we modified a conventional reverse transcription polymerase chain reaction assay for the specific detection of EHDV. This modified assay could detect various strains of EHDV isolated in Japan, Australia, and North America. Furthermore, the assay permitted the detection of EHDV-7 in blood cells collected from seven of the eight cattle. We believe that this modified assay will be a useful tool for the diagnosis of EHD.


Assuntos
Doenças dos Bovinos , Vírus da Doença Hemorrágica Epizoótica , Infecções por Reoviridae , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologia , Vírus da Doença Hemorrágica Epizoótica/genética , Japão/epidemiologia , Infecções por Reoviridae/diagnóstico , Infecções por Reoviridae/epidemiologia , Infecções por Reoviridae/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Transcrição Reversa , Sorogrupo
3.
Arch Virol ; 166(4): 1151-1156, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33547486

RESUMO

Tibet orbivirus (TIBOV) was initially isolated in Tibet in 2009 and subsequently in Guangdong, Hunan, and Yunnan, China. We document the first isolation of TIBOV outside of China: two TIBOV isolates from Culicoides collected in 2009 and 2010 in Kagoshima, Japan. Their complete genome sequences were also determined. Our results suggest that the two virus isolates are of novel serotypes, evident by variability within genome segment 2 encoding VP2. These new putative TIBOV serotypes will help with future virus surveillance and with the evaluation of its potential to cause disease in domestic ruminants.


Assuntos
Genoma Viral/genética , Orbivirus/genética , Orbivirus/isolamento & purificação , Animais , Ceratopogonidae/virologia , Genômica , Japão , Orbivirus/classificação , Filogenia , RNA Viral/genética , Homologia de Sequência , Sorogrupo , Proteínas Virais/genética
4.
Arch Virol ; 165(12): 2903-2908, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32894348

RESUMO

A novel orbivirus (genus Orbivirus, family Reoviridae), designated Yonaguni orbivirus (YONOV), was isolated from bovine blood collected on a subtropical island of Japan in 2015. The YONOV genome (20,054 nucleotides in total) has a coding arrangement similar to those of mosquito-borne orbiviruses. YONOV has a close genetic relationship to mosquito-borne orbiviruses, especially to Mobuck virus (MBV), which was isolated in North America. However, YONOV and MBV share less than 74% nucleotide sequence identity in the major subcore protein (T2) coding sequence, which satisfies the criterion for species demarcation. It is still uncertain whether YONOV should be assigned to a novel species in the genus Orbivirus.


Assuntos
Genoma Viral , Orbivirus/classificação , Orbivirus/isolamento & purificação , Filogenia , Infecções por Reoviridae/veterinária , Proteínas Virais/genética , Animais , Bovinos/virologia , Culicidae/virologia , Japão , Fases de Leitura Aberta , Infecções por Reoviridae/virologia , Análise de Sequência de DNA
5.
Microbiol Resour Announc ; 9(39)2020 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-32972940

RESUMO

Akabane virus (AKAV) (genus Orthobunyavirus, family Peribunyaviridae) is an arthropod-borne virus that causes congenital abnormalities in ruminants. Here, we report the complete genome sequences of two AKAV strains causing nonsuppurative encephalomyelitis in cattle by postnatal infection in Japan.

6.
J Vet Med Sci ; 81(10): 1496-1503, 2019 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-31447461

RESUMO

A surveillance of Culicoides biting midges with light suction traps was conducted in the northern region of Honshu, main island of Japan, during the summers and autumns of 2009 and 2010. A total of 106 trap collections across 37 cattle farms were investigated for the structure and distribution of Culicoides species. Forty-thousand and one hundred forty-nine specimens of Culicoides biting midges were identified at the species level, and ≥19 species were included in the specimens. Culicoides oxystoma, which is a known major vector of Akabane virus (AKAV), appeared not to have expanded in northern Honshu during the surveillance. Of the potential AKAV vectors suggested by a previous laboratory experiment, C. tainanus and C. punctatus widely infested cowsheds across northern Honshu. The AKAV circulation was confirmed by serological surveillance of sentinel cattle in northern Honshu during the summer and autumn of 2010 and, consequently, >200 calves affected by the virus were identified as of spring 2011. Our surveillance demonstrated that C. tainanus and C. punctatus were widely spread and often dominated at cattle farms in/around the seroconverted regions, and our results thus suggest that these species played a critical role in the AKAV transmission in 2010. Because the distribution ranges of C. tainanus and C. punctatus cover almost all of mainland Japan, a potential risk of AKAV transmission might be expected even in areas outside the range of C. oxystoma.


Assuntos
Doenças dos Bovinos/transmissão , Doenças dos Bovinos/virologia , Ceratopogonidae/virologia , Insetos Vetores/virologia , Orthobunyavirus , Animais , Bovinos , Surtos de Doenças , Japão/epidemiologia , Testes Sorológicos
7.
J Med Entomol ; 56(2): 533-539, 2019 02 25.
Artigo em Inglês | MEDLINE | ID: mdl-30418597

RESUMO

To test their virus susceptibility and capacity as transmission vectors, Japanese Culicoides species were artificially fed a mixture of Akabane virus (AKAV) and bovine blood, and virus recovery was attempted from infected midges 9-11 d post-exposure. Culicoides tainanus, C. punctatus, C. humeralis, C. jacobsoni, C. oxystoma, and C. asiana were found to be orally susceptible to AKAV. Virus titers in single infected midges of C. tainanus, C. oxystoma, C. punctatus, and C. jacobsoni ranged from 100.75 to 104.0 TCID50 (tissue culture infectious dose). The titers in the infected C. oxystoma were significantly higher than those in the other infected species. Viral RNA was detected from both midges testing positive and those testing negative for infectious virus particles, but the viral RNA copies in the infectious virus-negative midges were significantly lower than those in the infectious virus-positive midges. Lower viral amplification, limited dissemination or both caused by tissue barriers might occur in infected midges from which infectious viruses were undetectable. A fully disseminated infection was developed in orally infected C. oxystoma and C. tainanus. This finding indicates their capacity to transmit AKAV, assuming that salivary gland barriers have limited effects on viral entry to and replication in salivary gland tissue. This result also suggests that the other orally susceptible species are potentially competent for AKAV transmission and would be considered active vectors of its spread.


Assuntos
Ceratopogonidae/virologia , Insetos Vetores/virologia , Orthobunyavirus/fisiologia , Animais , Comportamento Alimentar
8.
J Vet Diagn Invest ; 30(6): 855-861, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30204057

RESUMO

Peaton virus (PEAV; family Peribunyaviridae, genus Orthobunyavirus) appears to be capable of producing congenital malformations in ruminants; however, its pathogenicity remains unknown given its relatively low incidence. We evaluated the relationship between congenital abnormalities of calves and PEAV infection by serologic, epidemiologic, pathologic, and virologic investigations using specimens from 31 malformed calves in the years 1996-2016 in Japan. Antibody testing was carried out for known teratogenic viruses, including Akabane, Aino, Chuzan, and bovine viral diarrhea viruses, in the precolostral sera of these abnormal calves, but all results were negative. However, all 31 malformed calves were positive for antibodies against PEAV. A PEAV-specific gene was amplified from central nervous system tissues from a stillborn calf delivered in April 2007, and its nucleotide sequence was identical with that of PEAV isolated from healthy sentinel cattle in September 2006. These findings indicate that PEAV can cause bovine congenital anomalies.


Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/patologia , Bovinos/anormalidades , Anormalidades Congênitas/veterinária , Orthobunyavirus/fisiologia , Animais , Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/patologia , Infecções por Bunyaviridae/virologia , Doenças dos Bovinos/virologia , Anormalidades Congênitas/patologia , Anormalidades Congênitas/virologia , Japão
9.
Virus Genes ; 54(5): 729-732, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30069670

RESUMO

Two virus strains, tentatively designated as ON-6/P/05 and ON-7/E/05, were isolated from blood samples of healthy cattle in the Yaeyama Islands, located in the southwestern-most region of Japan, in 2005. Ultrastructural observations of infected baby hamster (BHK-21) cells revealed that the viruses had features consistent with those of orbivirus. As with other orbiviruses, the viral genome consists of 10 double-stranded RNA segments. The full genome sequence of ON-6/P/05 was determined and shared high nucleotide and amino acid identities (90.07-98.22% nucleotide identity; 96.16-99.72% amino acid identity) with that of Sathuvachari virus (SVIV), a member of the species Sathuvachari virus of the genus Orbivirus, originally isolated from starlings collected in southern India in 1963. The sequence of segment two of ON-7/E/05 was identical to that of ON-6/P/05. The isolation of SVIV from cattle also indicated that the virus has a wider host range than previously thought. The potential pathogenicity of SVIV in domestic animals should be considered in future disease surveillance within its distribution range.


Assuntos
Bovinos/virologia , Genoma Viral , Orbivirus/genética , Orbivirus/isolamento & purificação , Animais , Linhagem Celular , Cricetinae , Feminino , Japão , Tipagem Molecular , Orbivirus/classificação , Especificidade da Espécie
10.
Emerg Infect Dis ; 24(5): 902-905, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29664367

RESUMO

During October-December 2015, an epizootic hemorrhagic disease outbreak occurred in cattle in Japan. Forty-six animals displayed fever, anorexia, cessation of rumination, salivation, and dysphagia. Virologic, serologic, and pathologic investigations revealed the causative agent was epizootic hemorrhagic disease virus serotype 6. Further virus characterization is needed to determine virus pathogenicity.


Assuntos
Doenças dos Bovinos/virologia , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Infecções por Reoviridae/veterinária , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Surtos de Doenças , Vírus da Doença Hemorrágica Epizoótica/genética , Japão/epidemiologia , Filogenia , Infecções por Reoviridae/epidemiologia , Infecções por Reoviridae/virologia
11.
Microbes Environ ; 33(1): 77-82, 2018 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-29491247

RESUMO

Salmonella and Campylobacter cause foodborne enteritis mainly via the consumption of raw/undercooked contaminated poultry meat and products. Broiler flocks are primarily colonized with these bacteria; however, the underlying etiology remains unclear. The present study was conducted in order to obtain further information on the prevalence and genotypic distribution of Salmonella and Campylobacter in free-living crows and broiler flocks in a region for 2 years, thereby facilitating estimations of the potential risk of transmission of C. jejuni from crows to broiler flocks. Salmonella serovars Bredeney and Derby were isolated from 8 and 3 out of 123 captured crows, respectively, both of which are not common in broiler chickens. Campylobacter were isolated from all 89 crows tested and C. jejuni was prevalent (85 crows). Pulsed field gel electrophoresis showed broad diversity in the crow isolates of C. jejuni. However, 3 crow isolates and 2 broiler isolates showing similar banding patterns were assigned to different sequence types in multi-locus sequence typing. These results indicate that crows do not share Salmonella serovars with broilers, and harbor various genotypes of C. jejuni that differ from those of broiler flocks. Thus, our results indicate that crows are not a potential vector of these bacteria to broiler flocks in this region.


Assuntos
Campylobacter/isolamento & purificação , Galinhas/microbiologia , Corvos/microbiologia , Salmonella/isolamento & purificação , Animais , Campylobacter/classificação , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/veterinária , Eletroforese em Gel de Campo Pulsado , Genótipo , Japão/epidemiologia , Tipagem de Sequências Multilocus , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Aves Domésticas/microbiologia , Prevalência , Salmonella/classificação , Salmonelose Animal/epidemiologia , Sorogrupo
12.
Infect Genet Evol ; 53: 38-46, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28506840

RESUMO

We characterized genome segments 2, 3 and 6 (Seg-2, Seg-3 and Seg-6) of 11 Japanese strains of epizootic hemorrhagic disease (EHD) virus (EHDV) isolated in 1985-2013. The Japanese strains were divisible into two groups based on phylogenetic analyses of the nucleotide sequences of Seg-2 and Seg-6. In both of the phylogenetic trees based on Seg-2 and Seg-6, seven of the 11 Japanese strains were grouped together with EHDV-2 and EHDV-7 strains, and the other four Japanese strains were grouped with EHDV-1 strains. The phylogenetic analysis of Seg-2 among EHDV strains identified 10 of the 11 Japanese strains as EHDV-1, EHDV-2 or EHDV-7. The other Japanese strain, ON-4/B/98, isolated from an asymptomatic cow in 1998 was in the same group as the EHDV-2 and EHDV-7 strains in the phylogenetic trees based on Seg-2 and Seg-6, but the results suggested that the strain belongs to another serotype. We thus conducted a serum neutralization test to identify that serotype by using anti-EHDV-2 and anti-EHDV-7 rabbit sera. We observed that the ON-4/B/98 strain was not sufficiently neutralized by any of the antisera, which suggests that the strain could be assigned into a new serotype, tentatively named 'EHDV-10.' Sequences of Seg-3 were also determined, and all of the Japanese strains were grouped together with Australian strains, suggesting that the Japanese strains are a part of EHDV distributed in the Asia-Pacific region. The data obtained herein would be beneficial for the diagnosis and prevention of EHD in Japan and neighboring countries.


Assuntos
Doenças dos Bovinos/epidemiologia , Genoma Viral , Vírus da Doença Hemorrágica Epizoótica/genética , Filogenia , RNA Viral/genética , Infecções por Reoviridae/veterinária , Animais , Bovinos , Doenças dos Bovinos/virologia , Genótipo , Vírus da Doença Hemorrágica Epizoótica/classificação , Vírus da Doença Hemorrágica Epizoótica/imunologia , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Soros Imunes , Japão/epidemiologia , Tipagem Molecular , Testes de Neutralização , Filogeografia , Coelhos , Infecções por Reoviridae/epidemiologia , Infecções por Reoviridae/virologia , Sorogrupo
14.
J Vet Med Sci ; 79(5): 904-911, 2017 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-28392506

RESUMO

In September and October 2015, suspected cases of bovine ephemeral fever (BEF) were reported in the mainland region of Kagoshima Prefecture and on Tanegashima Island. The genome of the BEF virus (BEFV) was detected in the diseased cows and the cows that had recovered. The serum obtained from the affected cows contained high titers of BEFV-neutralizing antibody. In total, 18 affected cows were demonstrated to be infected with BEFV during the outbreak. Our findings showed evidence that BEF occurred in mainland Japan after a 23-year absence. Phylogenetic analysis based on the surface glycoprotein (G) gene revealed that BEFVs detected in the affected cows were genetically distinct from previous Japanese BEFVs, but were close to BEFVs circulating in Taiwan and mainland China in recent years. Amino acid substitution in the neutralizing epitope domains of the G protein was limited between the detected viruses and the vaccine strain (YHL isolate), and high titers of the neutralizing antibody against the YHL isolate were induced in the infected cattle during the disease occurrences. Therefore, current BEF vaccines probably elicit protective immunity against the BEFVs detected in 2015, although their effectiveness should be assessed. Since the BEFV vaccination rates are estimated to be low, a BEF outbreak should be considered a possibility in mainland Japan.


Assuntos
Febre Efêmera/diagnóstico , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Bovinos , Surtos de Doenças/prevenção & controle , Surtos de Doenças/veterinária , Febre Efêmera/epidemiologia , Febre Efêmera/prevenção & controle , Febre Efêmera/virologia , Vírus da Febre Efêmera Bovina/classificação , Vírus da Febre Efêmera Bovina/genética , Vírus da Febre Efêmera Bovina/isolamento & purificação , Feminino , Insetos Vetores/virologia , Japão/epidemiologia , Filogenia , Vacinas Virais/administração & dosagem
15.
J Vet Med Sci ; 79(4): 774-779, 2017 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-28302930

RESUMO

We isolated an arbovirus from bovine blood in Indonesia. The arbovirus was obtained from the plasma of a cow showing no clinical symptoms in West Java in February 2014, and was identified as Akabane virus (AKAV) by AKAV-specific RT-PCR and subsequent sequence analysis. Phylogenetic analysis based on partial S segment indicated the AKAV isolate, WJ-1SA/P/2014, was most closely related with two isolates from Israel and Turkey reported in 2001 and 2015, respectively, and that WJ-1SA/P/2014 isolate belongs to AKAV genogroup Ib. This is the first isolation of AKAV from Indonesia.


Assuntos
Arbovírus/genética , Bovinos/virologia , Orthobunyavirus/genética , Animais , Arbovírus/classificação , Arbovírus/isolamento & purificação , Células Cultivadas , Indonésia , Orthobunyavirus/classificação , Orthobunyavirus/isolamento & purificação
16.
BMC Vet Res ; 12(1): 125, 2016 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-27342576

RESUMO

BACKGROUND: In Japan, epizootic arboviral infections have severely impacted the livestock industry for a long period. Akabane, Aino, Chuzan, bovine ephemeral fever and Ibaraki viruses have repeatedly caused epizootic abnormal births and febrile illness in the cattle population. In addition, Peaton, Sathuperi, Shamonda and D'Aguilar viruses and epizootic hemorrhagic virus serotype 7 have recently emerged in Japan and are also considered to be involved in abnormal births in cattle. The above-mentioned viruses are hypothesized to circulate in tropical and subtropical Asia year round and to be introduced to temperate East Asia by long-distance aerial dispersal of infected vectors. To watch for arbovirus incursion and assess the possibility of its early warning, monitoring for arboviruses was conducted in the Yaeyama Islands, located at the most southwestern area of Japan, between 1994 and 2014. RESULTS: Blood sampling was conducted once a year, in the autumn, in 40 to 60 healthy cattle from the Yaeyama Islands. Blood samples were tested for arboviruses. A total of 33 arboviruses including Akabane, Peaton, Chuzan, D' Aguilar, Bunyip Creek, Batai and epizootic hemorrhagic viruses were isolated from bovine blood samples. Serological surveillance for the bovine arboviruses associated with cattle diseases in young cattle (ages 6-12 months: had only been alive for one summer) clearly showed their frequent incursion into the Yaeyama Islands. In some cases, the arbovirus incursions could be detected in the Yaeyama Islands prior to their spread to mainland Japan. CONCLUSIONS: We showed that long-term surveillance in the Yaeyama Islands could estimate the activity of bovine arboviruses in neighboring regions and may provide a useful early warning for likely arbovirus infections in Japan. The findings in this study could contribute to the planning of prevention and control for bovine arbovirus infections in Japan and cooperative efforts among neighboring countries in East Asia.


Assuntos
Infecções por Arbovirus/veterinária , Doenças dos Bovinos/epidemiologia , Animais , Anticorpos Antivirais/sangue , Infecções por Arbovirus/sangue , Infecções por Arbovirus/epidemiologia , Infecções por Arbovirus/prevenção & controle , Arbovírus/isolamento & purificação , Bovinos , Doenças dos Bovinos/prevenção & controle , Ilhas , Japão , Vigilância da População
17.
J Vet Med Sci ; 78(3): 447-50, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26498401

RESUMO

A serological survey of caprine arthritis-encephalitis virus (CAEV) infection was conducted from September 2006 to February 2007 in Japan. A total of 857 serum samples were collected from 113 herds in 28 prefectures and were analyzed for the presence of CAEV antibodies using agar gel immunodiffusion test. The seroprevalence of CAEV infection at the herd and animal levels was 15.0% (17/113) and 10.0% (86/857), respectively. Large farms with more than 10 goats and with animals for dairy and breeding purposes had higher seroprevalence (P<0.05). The results of this study provide useful information to consider effective control programs against CAEV infection in Japan.


Assuntos
Vírus da Artrite-Encefalite Caprina , Doenças das Cabras/epidemiologia , Infecções por Lentivirus/veterinária , Animais , Feminino , Doenças das Cabras/virologia , Cabras , Japão/epidemiologia , Infecções por Lentivirus/epidemiologia , Masculino , Prevalência
18.
J Virol Methods ; 225: 9-15, 2015 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-26341063

RESUMO

TaqMan assays were developed for the broad-range detection of arboviruses belonging to Simbu serogroup lineage 1 in the genus Orthobunyavirus and also for the specific detection of three viruses in the lineage, Akabane, Aino and Peaton viruses (AKAV, AINOV and PEAV, respectively). A primer and probe set was designed for the broad-range detection of Simbu serogroup lineage 1 (Pan-Simbu1 set) mainly targeting AKAV, AINOV, PEAV, Sathuperi and Shamonda viruses (SATV and SHAV), and the forward and reverse primers of the Pan-Simbu1 set were also used for the specific detection of AKAV with another probe (AKAV-specific set). In addition, two more primer and probe sets were designed for AINOV- and PEAV-specific detection, respectively (AINOV- and PEAV-specific sets). All of the four primer and probe sets successfully detected targeted viruses, and thus broad-range and specific detection of all the targeted viruses can be achieved by using two multiplex assays and a single assay in a dual (two-color) assay format when another primer and probe set for a bovine ß-actin control is also used. The assays had an analytical sensitivity of 10 copies/tube for AKAV, at least 100 copies/tube for AINOV, 100 copies/tube for PEAV, one copy/tube for SATV and at least 10 copies/tube for SHAV, respectively. Diagnostic sensitivity of the assays was tested with field-collected bovine samples, and the results suggested that the sensitivity was higher than that of a conventional RT-PCR. These data indicate that the newly developed TaqMan assays will be useful tools for the diagnosis and screening of field-collected samples for infections of AKAV and several other arboviruses belonging to the Simbu serogroup lineage 1.


Assuntos
Arbovírus/classificação , Arbovírus/isolamento & purificação , Infecções por Bunyaviridae/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Orthobunyavirus/classificação , Orthobunyavirus/isolamento & purificação , Virologia/métodos , Animais , Arbovírus/genética , Infecções por Bunyaviridae/veterinária , Infecções por Bunyaviridae/virologia , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/virologia , Primers do DNA/genética , Sondas de Oligonucleotídeos/genética , Orthobunyavirus/genética , Sensibilidade e Especificidade
19.
J Vet Med Sci ; 77(10): 1253-9, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26018356

RESUMO

In Japan in 2013, two cattle in the northwestern part of Kagoshima Prefecture developed fever and swallowing difficulty and were suspected of having Ibaraki disease. The epizootic hemorrhagic virus (EHDV) genome was detected from diseased and asymptomatic cattle by reverse transcription-polymerase chain reaction (RT-PCR). High neutralization antibody titers to Ibaraki virus (IBAV) ranging from 1:128 to 1:1,024 were observed in the RT-PCR-positive cattle, and the virus was isolated in one of the IBAV-positive farms. A pairwise alignment and phylogenetic analysis based on the major outer coat protein VP2 encoded in segment 2 revealed a close relationship between the isolated viruses and previous IBAV isolates. The phylogeny of VP2 also suggested that an IBAV variant isolated in 1997 was distinct from IBAV and sorted into a heterogeneous serotype, EHDV serotype 7. The findings revealed the reemergence of Ibaraki disease in Japan after a 26-year absence. Interestingly, the co-circulation of EHDV serotype 1 with IBAV was observed in the affected region, suggesting the potential reassortment between two heterogeneous serotypes in the field. Sentinel surveillance in Kagoshima Prefecture indicated that the incursion of IBAV occurred in October 2013 and that its spread was limited within the small area. Inadequate environmental temperatures for vector transmission in late autumn might have limited the virus spread to a wider region. The reemergence of Ibaraki disease showed us the importance of continuous vaccination to prevent economic losses.


Assuntos
Doenças dos Bovinos/virologia , Vírus da Doença Hemorrágica Epizoótica/isolamento & purificação , Infecções por Reoviridae/veterinária , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Vírus da Doença Hemorrágica Epizoótica/genética , Japão/epidemiologia , Filogenia , Infecções por Reoviridae/epidemiologia , Infecções por Reoviridae/virologia , Vigilância de Evento Sentinela
20.
J Vet Med Sci ; 77(4): 455-60, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25648276

RESUMO

In September 2012, several cows and a calf showed decreased activity, anorexia and fever on Ishigaki Island, Okinawa Prefecture, Japan, and the cases were diagnosed as bovine ephemeral fever (BEF). We isolated BEF virus (BEFV) from one of the affected cows and then determined the complete genome sequence of the G gene, which encodes a class I transmembrane glycoprotein of BEFV. The BEFV isolate in this case, ON-3/E/12, was sorted into the same cluster as other BEFV isolates in Japan, Taiwan and China obtained in 1996-2004 and was most closely related to a 2002 Chinese isolate, JT02L, according to the phylogenetic analysis of the complete G gene. Since inactivated vaccines for BEF available in Japan are considered effective against the ON-3/E/12 isolate as well as other isolates in East Asia from 1996-2004, annual vaccination should be conducted to prevent BEF in Okinawa. Additionally, in this study, we developed an RT-PCR assay to detect the BEFV gene in Japan and neighboring countries. Our assay was able to amplify target sequences in all of the tested BEFV isolates, including 18 isolates in Japan and another isolate in Australia. The assay was found to be useful also for testing RNA samples extracted from bovine peripheral blood mononuclear cells, and the detection limit of the assay was 10 copies per tube. We believe that our assay would be an important tool for the screening of BEFV infection and the diagnosis of BEF.


Assuntos
Vírus da Febre Efêmera Bovina/isolamento & purificação , Febre Efêmera/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Bovinos , Febre Efêmera/epidemiologia , Febre Efêmera/virologia , Vírus da Febre Efêmera Bovina/genética , Feminino , Regulação Viral da Expressão Gênica/fisiologia , Japão , Filogenia , Proteínas Virais/genética , Proteínas Virais/metabolismo
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