RESUMO
Myxomatosis is a recurrent problem on rabbit farms throughout Europe despite the success of vaccines. To identify gene variations of field and vaccine strains that may be responsible for changes in virulence, immunomodulation, and immunoprotection, the genomes of 6 myxoma virus (MYXV) strains were sequenced: German field isolates Munich-1, FLI-H, 2604, and 3207; vaccine strain MAV; and challenge strain ZA. The analyzed genomes ranged from 147.6 kb (strain MAV) to 161.8 kb (strain 3207). All sequences were affected by several mutations, covering 24 to 93 open reading frames (ORFs) and resulted in amino acid substitutions, insertions, or deletions. Only strains Munich-1 and MAV revealed the deletion of 10 ORFs (M007L to M015L) and 11 ORFs (M007L to M008.1L and M149R to M008.1R), respectively. Major differences were observed in the 27 immunomodulatory proteins encoded by MYXV. Compared to the reference strain Lausanne, strains FLI-H, 2604, 3207, and ZA showed the highest amino acid identity (>98.4%). In strains Munich-1 and MAV, deletion of 5 and 10 ORFs, respectively, was observed, encoding immunomodulatory proteins with ankyrin repeats or members of the family of serine protease inhibitors. Furthermore, putative immunodominant surface proteins with homology to vaccinia virus (VACV) were investigated in the sequenced strains. Only strain MAV revealed above-average frequencies of amino acid substitutions and frameshift mutations. Finally, we performed recombination analysis and found signs of recombination in vaccine strain MAV. Phylogenetic analysis showed a close relationship of strain MAV and the MSW strain of Californian MYXV. However, in a challenge model, strain MAV provided full protection against lethal challenges with strain ZA. IMPORTANCE: Myxoma virus (MYXV) is pathogenic for European rabbits and two North American species. Due to sophisticated strategies in immune evasion and oncolysis, MYXV is an important model virus for immunological and pathological research. In its natural hosts, MYXV causes a benign infection, whereas in European rabbits, it causes the lethal disease myxomatosis. Since the introduction of MYXV into Australia and Europe for the biological control of European rabbits in the 1950s, a coevolution of host and pathogen has started, selecting for attenuated virus strains and increased resistance in rabbits. Evolution of viruses is a continuous process and influences the protective potential of vaccines. In our analyses, we sequenced 6 MYXV field, challenge, and vaccine strains. We focused on genes encoding proteins involved in virulence, host range, immunomodulation, and envelope composition. Genes affected most by mutations play a role in immunomodulation. However, attenuation cannot be linked to individual mutations or gene disruptions.
Assuntos
Variação Genética , Genoma Viral , Myxoma virus/genética , Infecções por Poxviridae/virologia , Substituição de Aminoácidos , Animais , Repetição de Anquirina , Apoptose , Linhagem Celular , Chlorocebus aethiops , Evolução Molecular , Genômica/métodos , Imunomodulação , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/virologia , Leucócitos/imunologia , Leucócitos/metabolismo , Mutação , Myxoma virus/classificação , Myxoma virus/imunologia , Fases de Leitura Aberta , Filogenia , Infecções por Poxviridae/imunologia , Infecções por Poxviridae/prevenção & controle , Ligação Proteica , Mapeamento de Interação de Proteínas , Coelhos , Receptores Imunológicos , Proteínas Virais/genética , Proteínas Virais/imunologia , Proteínas Virais/metabolismo , Vacinas Virais/genética , Vacinas Virais/imunologiaRESUMO
Orthobunyaviruses are enveloped viruses that are arthropod-transmitted and cause disease in humans and livestock. Viral attachment and entry are mediated by the envelope glycoproteins Gn and Gc, and the major glycoprotein, Gc, of certain orthobunyaviruses is targeted by neutralizing antibodies. The domains in which the epitopes of such antibodies are located on the glycoproteins of the animal orthobunyavirus Schmallenberg virus (SBV) have not been identified. Here, we analysed the reactivity of a set of mAbs and antisera against recombinant SBV glycoproteins. The M-segment-encoded proteins Gn and Gc of SBV were expressed as full-length proteins, and Gc was also produced as two truncated forms, which consisted of its amino-terminal third and carboxyl-terminal two-thirds. The sera from convalescent animals reacted only against the full-length Gc and its subdomains and not against the SBV glycoprotein Gn. Interestingly, the amino-terminal domain of SBV-Gc was targeted not only by polyclonal sera but also by the majority of murine mAbs with a neutralizing activity. Furthermore, the newly defined amino-terminal domain of about 230 aa of the SBV Gc protein could be affinity-purified and further characterized. This major neutralizing domain might be relevant for the development of prophylactic, diagnostic and therapeutic approaches for SBV and other orthobunyaviruses.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Infecções por Bunyaviridae/imunologia , Orthobunyavirus/imunologia , Proteínas do Envelope Viral/química , Proteínas do Envelope Viral/imunologia , Motivos de Aminoácidos , Animais , Infecções por Bunyaviridae/virologia , Humanos , Imunidade Humoral , Camundongos , Orthobunyavirus/química , Orthobunyavirus/genética , Estrutura Terciária de Proteína , Proteínas do Envelope Viral/genéticaRESUMO
Since 2013, highly virulent porcine epidemic diarrhea virus has caused considerable economic losses in the United States. To determine the relation of US strains to those recently causing disease in Germany, we compared genomes and found that the strain from Germany is closely related to variants in the United States.
Assuntos
Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína/genética , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/virologia , Animais , Genoma Viral , Alemanha/epidemiologia , Filogenia , Vírus da Diarreia Epidêmica Suína/ultraestrutura , Análise de Sequência de DNA , Suínos , Estados Unidos/epidemiologia , Proteínas Virais/genéticaRESUMO
UNLABELLED: In February 2013, very severe acute clinical symptoms were observed in calves, heifers, and dairy cattle in several farms in North Rhine Westphalia and Lower Saxony, Germany. Deep sequencing revealed the coexistence of three distinct genome variants within recent highly virulent bovine viral diarrhea virus type 2 (BVDV-2) isolates. While the major portion (ca. 95%) of the population harbored a duplication of a 222-nucleotide (nt) segment within the p7-NS2-encoding region, the minority reflected the standard structure of a BVDV-2 genome. Additionally, unusual mutations were found in both variants, within the highly conserved p7 protein and close to the p7-NS2 cleavage site. Using a reverse genetic system with a BVDV-2a strain harboring a similar duplication, it could be demonstrated that during replication, genomes without duplication are generated de novo from genomes with duplication. The major variant with duplication is compulsorily escorted by the minor variant without duplication. RNA secondary structure prediction allowed the analysis of the unique but stable mixture of three BVDV variants and also provided the explanation for their generation. Finally, our results suggest that the variant with duplication plays the major role in the highly virulent phenotype. IMPORTANCE: This study emphasizes the importance of full-genome deep sequencing in combination with manual in-depth data analysis for the investigation of viruses in basic research and diagnostics. Here we investigated recent highly virulent bovine viral diarrhea virus isolates from a 2013 series of outbreaks. We discovered a unique special feature of the viral genome, an unstable duplication of 222 nucleotides which is eventually deleted by viral polymerase activity, leading to an unexpectedly mixed population of viral genomes for all investigated isolates. Our study is of high importance to the field because we demonstrate that these insertion/deletion events allow another level of genome plasticity of plus-strand RNA viruses, in addition to the well-known polymerase-induced single nucleotide variations which are generally considered the main basis for viral adaptation and evolution.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina Tipo 2/genética , Vírus da Diarreia Viral Bovina Tipo 2/isolamento & purificação , Animais , Bovinos , Vírus da Diarreia Viral Bovina Tipo 2/classificação , Vírus da Diarreia Viral Bovina Tipo 2/patogenicidade , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Filogenia , RNA Viral/química , RNA Viral/genética , Sequências Repetitivas de Ácido Nucleico , VirulênciaRESUMO
Bungowannah virus is the most divergent pestivirus, and both origin and reservoir host have not been identified so far. We therefore performed in vitro tropism studies, which showed that Bungowannah virus differs remarkably from other pestiviruses. Interestingly, cell lines of vervet monkey, mouse, human and even of bat origin were susceptible. This broad in vitro tropism was not observed for a chimeric bovine viral diarrhoea virus (BVDV) expressing all structural proteins of Bungowannah virus. The viral envelope was not sufficient to completely transfer the cell tropism of Bungowannah virus to another pestivirus, and viral RNA replication was either markedly reduced or not detectable in a number of different cell lines for the tested BVDV strain and the chimera. We therefore suggest that the replication machinery together with the viral envelope is responsible for the unique broad cell tropism of Bungowannah virus.
Assuntos
Pestivirus/fisiologia , Proteínas do Envelope Viral/metabolismo , Tropismo Viral , Animais , Linhagem Celular , Quirópteros , Chlorocebus aethiops , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/fisiologia , Humanos , Camundongos , Pestivirus/genética , Proteínas do Envelope Viral/genéticaRESUMO
BACKGROUND: Schmallenberg virus (SBV), a novel orthobunyavirus of the Simbu serogroup, was first identified in October 2011 in dairy cattle in Germany, where it caused fever, diarrhea and a drop in milk yield. Since then, SBV additionally has been detected in adult sheep and goats. Although symptoms of acute infection were not observed, infection during a vulnerable phase of pregnancy caused congenital malformations and stillbirths. In view of the current situation and the possible emergence of further Simbu serogroup members, a pan-Simbu real-time reverse transcriptase (RT) PCR system for the reliable detection of Simbu serogroup viruses should be developed. METHODS: In this study a pan-Simbu real-time RT-PCR system was established and compared to several SBV real-time RT-PCR assays. All PCR-systems were tested using a panel of different Simbu serogroup viruses as well as several field samples from diseased cattle, sheep and goats originating from all over Germany. Several pan-Simbu real-time RT-PCR products were sequenced via Sanger sequencing. Furthermore, in silico analyses were performed to investigate suitability for the detection of further orthobunyaviruses. RESULTS: All tested members of the Simbu serogroup (n = 14) as well as most of the field samples were successfully detected by the pan-Simbu real-time RT-PCR system. The comparison of this intercalating dye assay with different TaqMan probe-based assays developed for SBV diagnostics confirmed the functionality of the pan-Simbu assay for screening purposes. However, the SBV-TaqMan-assay SBV-S3 delivered the highest analytical sensitivity of less than ten copies per reaction for duplex systems including an internal control. In addition, for confirmation of SBV-genome detection the highly specific SBV-M1 assay was established. CONCLUSION: The pan-Simbu real-time RT-PCR system was able to detect all tested members of the Simbu serogroup, most of the SBV field samples as well as three tested Bunyamwera serogroup viruses with a suitable sensitivity. According to in silico analyses, this system seems to be able to detect a broad orthobunyavirus spectrum. As an additional feature of the pan-Simbu real-time RT-PCR system, subsequent species classification via sequencing is feasible. Regarding SBV diagnostics, the performance of the S-segment targeting SBV-S3 assay was superior with respect to the analytical sensitivity.
Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/diagnóstico , Doenças das Cabras/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Doenças dos Ovinos/diagnóstico , Vírus Simbu/isolamento & purificação , Animais , Infecções por Bunyaviridae/diagnóstico , Infecções por Bunyaviridae/virologia , Bovinos , Doenças dos Bovinos/virologia , Alemanha , Doenças das Cabras/virologia , Cabras , Técnicas de Diagnóstico Molecular/métodos , Sensibilidade e Especificidade , Ovinos , Doenças dos Ovinos/virologia , Vírus Simbu/genética , Fatores de TempoRESUMO
Schmallenberg virus (SBV), an orthobunyavirus of the Simbu serogroup, recently emerged in Europe and has been suggested to be a Shamonda/Sathuperi virus reassortant. Results of full-genome and serologic investigations indicate that SBV belongs to the species Sathuperi virus and is a possible ancestor of the reassortant Shamonda virus.
Assuntos
Evolução Molecular , Orthobunyavirus/genética , Vírus Reordenados/genética , Animais , Infecções por Bunyaviridae/veterinária , Infecções por Bunyaviridae/virologia , Linhagem Celular , Genoma Viral , Orthobunyavirus/classificação , Filogenia , Recombinação Genética , Vírus Simbu/classificação , Vírus Simbu/genética , Especificidade da EspécieRESUMO
Schmallenberg virus was detected in cattle and sheep in northwestern Europe in 2011. To determine whether wild ruminants are also susceptible, we measured antibody seroprevalence in cervids (roe deer and red deer) in Belgium in 2010 and 2011. Findings indicated rapid spread among these deer since virus emergence ≈250 km away.
Assuntos
Infecções por Bunyaviridae/veterinária , Cervos/virologia , Orthobunyavirus/isolamento & purificação , Animais , Anticorpos Antivirais/sangue , Bélgica , Infecções por Bunyaviridae/história , História do Século XXI , Imunoglobulina G/sangue , Orthobunyavirus/imunologia , Estações do Ano , Estudos Soroepidemiológicos , Proteínas Virais/imunologiaRESUMO
In 2011, an unidentified disease in cattle was reported in Germany and the Netherlands. Clinical signs included fever, decreased milk production, and diarrhea. Metagenomic analysis identified a novel orthobunyavirus, which subsequently was isolated from blood of affected animals. Surveillance was initiated to test malformed newborn animals in the affected region.
Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/virologia , Surtos de Doenças/veterinária , Orthobunyavirus/isolamento & purificação , Animais , Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/virologia , Bovinos , Doenças dos Bovinos/epidemiologia , Linhagem Celular , Cricetinae , Alemanha/epidemiologia , Países Baixos/epidemiologia , Proteínas do Nucleocapsídeo/genética , Orthobunyavirus/classificação , Orthobunyavirus/genética , Filogenia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
A serological survey was conducted in 2007 in the breeding goat population in Poland to gain insights into the epidemiology of pestivirus infection. All breeding herds were included in the study and representative serum samples were taken in each herd to evaluate herd-level seroprevalence at 10% expected individual-level prevalence and 95% level of confidence. Altogether 1060 serum samples from 49 herds were tested with blocking ELISA and then the positive and inconclusive results were confirmed in a serum neutralisation test, which also allowed us to determine the pestivirus species responsible for seroconversion. Herd-level seroprevalence proved to be 10.2% and bovine viral diarrhoea virus type 1 (BVDV-1) was responsible for the seroconversion in seven out of eight cases. In the remaining serum sample the causative virus could not be identified due to a pronounced cross-neutralising activity possibly derived from multiple infections. This is the first report on the diagnosis of BVDV-1 infection in Polish goats.
Assuntos
Vírus da Diarreia Viral Bovina Tipo 1/isolamento & purificação , Doenças das Cabras/virologia , Infecções por Pestivirus/veterinária , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças das Cabras/epidemiologia , Cabras , Infecções por Pestivirus/epidemiologia , Infecções por Pestivirus/virologia , Polônia/epidemiologia , PrevalênciaRESUMO
Vaccines against rabbit haemorrhagic disease virus (RHDV) are commercially produced in experimentally infected rabbits. A genetically engineered and manufactured version of the major structural protein of RHDV (VP60) is considered to be an alternative approach for vaccine production. Plants have the potential to become an excellent recombinant production system, but the low expression level and insufficient immunogenic potency of plant-derived VP60 still hamper its practical use. In this study, we analysed the expression of a novel multimeric VP60-based antigen in four different plant species, including Nicotiana tabacum L., Solanum tuberosum L., Brassica napus L. and Pisum sativum L. Significant differences were detected in the expression patterns of the novel fusion antigen cholera toxin B subunit (CTB)::VP60 (ctbvp60(SEKDEL)) at the mRNA and protein levels. Pentameric CTB::VP60 molecules were only detected in N. tabacum and P. sativum, and displayed equal levels of CTB, at approximately 0.01% of total soluble protein (TSP), and traces of detectable VP60. However, strong enhancement of the CTB protein content via self-fertilization was only observed in P. sativum, where it reached up to 0.7% of TSP. In rabbits, a strong decrease in the protective vaccine dose required from 48-400 microg potato-derived VP60 [Castanon, S., Marin, M.S., Martin-Alonso, J.M., Boga, J.A., Casais, R., Humara, J.M., Ordas, R.J. and Parra, F. (1999) Immunization with potato plants expressing VP60 protein protects against rabbit hemorrhagic disease virus. J. Virol. 73, 4452-4455; Castanon, S., Martin-Alonso, J.M., Marin, M.S., Boga, J.A., Alonso, P., Parra, F. and Ordas, R.J. (2002) The effect of the promoter on expression of VP60 gene from rabbit hemorrhagic disease virus in potato plants. Plant Sci. 162, 87-95] to 0.56-0.28 microg antigenic VP60 (measured with VP60 enzyme-linked immunosorbent assay) of crude CTB::VP60 pea extracts was demonstrated. Rabbits immunized with pea-derived CTB::VP60 showed anti-VP60-specific antibodies, similar to RikaVacc((R))-immunized rabbits, and survived RHDV challenge.
Assuntos
Infecções por Caliciviridae/veterinária , Vírus da Doença Hemorrágica de Coelhos/imunologia , Pisum sativum/imunologia , Proteínas Estruturais Virais/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Infecções por Caliciviridae/prevenção & controle , Toxina da Cólera/imunologia , DNA de Plantas/genética , Ensaio de Imunoadsorção Enzimática , Expressão Gênica , Pisum sativum/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/imunologia , Regiões Promotoras Genéticas , Coelhos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Estruturais Virais/genéticaRESUMO
Bovine viral diarrhea (BVD) causes high economic losses in the cattle population worldwide. In Germany, an obligatory control program with detection and removal of persistently infected animals is in force since 2011. For molecular tracing of virus transmission, a comprehensive sequence data base of the currently circulating BVD viruses was established. Partial sequences of 1007 samples collected between 2008 and 2016 were generated. As dominant viruses, subtypes 1b (47.0%) and 1d (26.5%) could be identified with no marked geographic or sampling year effect, a much higher amount of BVDV-2c was detected in 2013 compared to other years, predominantly in Western Germany. In addition, subtypes 1a, 1e, 1f, 1h, 1g, 1k, and 2a were found. Interestingly, besides field-viruses, two different live-vaccine viruses were detected in tissue samples of newborn calves (n=37) whose mothers were immunized during pregnancy.
Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/epidemiologia , Vírus da Diarreia Viral Bovina/genética , Erradicação de Doenças/estatística & dados numéricos , Vacinas Virais/imunologia , Animais , Doença das Mucosas por Vírus da Diarreia Viral Bovina/prevenção & controle , Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Bovinos , Vírus da Diarreia Viral Bovina/classificação , Alemanha/epidemiologiaRESUMO
Bovine viral diarrhea (BVD) is one of the most important infectious diseases in cattle, causing major economic losses worldwide. Therefore, control programs have been implemented in several countries. In Germany, an obligatory nationwide eradication program has been in force since 2011. Its centerpiece is the detection of animals persistently infected (PI) with BVD virus, primarily based on the testing of ear tissue samples of all newborn calves for viral genome or antigen, and their removal from the cattle population. More than 48,000 PI animals have so far been detected and removed. Between the onset of the program and the end of 2016, the prevalence of these animals among all newborn calves decreased considerably, from 0.5% to less than 0.03%. The number of cattle holdings with PI animals likewise decreased from 3.44% in 2011 to only 0.16% in 2016. Since a large number of naïve, fully susceptible animals are now confronted with BVD virus, which is still present in the German cattle population, the challenge of the coming years will be the identification of remaining PI animals as quickly and efficiently as possible, and the efficient protection of BVD-free farms from reinfection.
RESUMO
Recombinant baculo viruses based on Autographa californica multiple nuclear polyhedrosis virus carrying vertebrate cell active expression cassettes, so-called BacMam viruses, are increasingly used as gene delivery vectors for vaccination of animals against pathogens. Different approaches for generation of BacMams exist and a variety of transfer vectors to improve target protein expression in vivo have been constructed. Here we describe a use of transfer vector which contains an insect cell-restricted expression cassette for the green fluorescent protein and thus enables easy monitoring of BacMam virus rescue, fast plaque purification of recombinants and their convenient titer determination and which has been proven to be efficacious for gene delivery in vaccination/challenge experiments.
Assuntos
Antígenos/imunologia , Baculoviridae/genética , Técnicas de Transferência de Genes , Nucleopoliedrovírus/imunologia , Vacinas/genética , Animais , Antígenos/genética , Baculoviridae/imunologia , Regulação Viral da Expressão Gênica , Vetores Genéticos , Proteínas de Fluorescência Verde , Mariposas/citologia , Mariposas/genética , Nucleopoliedrovírus/genética , Nucleopoliedrovírus/crescimento & desenvolvimento , Transdução Genética , Transfecção , Vacinas/imunologiaRESUMO
We detected Hepatitis E virus in serum samples of wild rabbits that were hunted in 1989 around the city of Greifswald, Germany. The recovery of one partial sequence and subsequent phylogenetic analysis indicates a close relationship to rabbit HEV sequences from France and suggests a long-established circulation of rabbit HEV in Europe.
Assuntos
Animais Selvagens/virologia , Sangue/virologia , Vírus da Hepatite E/isolamento & purificação , Coelhos/virologia , Animais , Animais Selvagens/sangue , Alemanha , Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Filogenia , Coelhos/sangueRESUMO
In November 2012, a dairy farmer in the district Kleve first observed a reduction in milk yield, respiratory symptoms, nasal discharge, fever, sporadic diarrhoea and sudden deaths in dairy cows and calves. In the following months, further farms were found infected with cattle showing similar clinical signs. An epidemiological investigation was carried out to identify the source of infection, the date of introduction, potential transmission pathways and to analyse the extent of the epidemic. Furthermore, laboratory analyses were conducted to characterise the causative agent. BVDV had been diagnosed in the index herd in December 2012, but due to the atypical clinical picture, the virus was not immediately recognised as the causative agent. Further laboratory analysis showed that this outbreak and subsequent infections in the area were caused by a BVD type 2c virus with a characteristic genome insertion, which seems to be associated with the occurrence of severe clinical symptoms in infected cattle. Epidemiological investigations showed that the probable date of introduction was in mid-October 2012. The high risk period was estimated as three months. A total of 21 affected farms with 5325 cattle were identified in two German Federal States. The virus was mainly transmitted by person contacts, but also by cattle trade and vehicles. The case-fatality rate was up to 60% and mortality in outbreak farms varied between 2.3 and 29.5%. The competent veterinary authorities imposed trade restrictions on affected farms. All persons who had been in contact with affected animals were advised to increase biosecurity measures (e.g. using farm-owned or disposable protective clothing). In some farms, affected animals were vaccinated against BVD to reduce clinical signs as an "emergency measure". These measures stopped the further spread of the disease.
RESUMO
Schmallenberg virus (SBV), an orthobunyavirus discovered in European livestock in late 2011 for the first time, causes premature or stillbirth and severe fetal malformation when cows and ewes are infected during pregnancy. Therefore, cattle of two holdings in the initially most affected area in Germany were closely monitored to describe the consequence for fetuses and newborn calves. Seventy-one calves whose mothers were naturally infected during the first five months of pregnancy were clinically, virologically, and serologically examined. One calve showed typical malformation, another one, born without visible abnormalities, was dead. Two cows aborted during the studied period; spleen and brain samples or meconium swabs were tested by real-time PCR, in none of the fetuses SBV-specific RNA was detectable and the tested fetal sera were negative in a commercially available antibody ELISA. In contrast, in nine clinically healthy calves high SBV-antibody titers were measurable before colostrum intake, and in meconium swabs of six of these animals viral RNA was present as well. The mothers of all nine seropositive calves were presumably infected between days 47 and 162 of gestation, which is within the critical timeframe for fetal infection suggested for SBV and related viruses.
Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/virologia , Animais , Infecções por Bunyaviridae/virologia , Bovinos , Feminino , Seguimentos , Gravidez , Reação em Cadeia da Polimerase em Tempo RealRESUMO
The complete genome sequence of the genotype 3 border disease virus strain Gifhorn has been determined; this strain was originally isolated from pigs. This represents the consensus sequence for the virus used to produce the bacterial artificial chromosome (BAC) cDNA clone pBeloGif3, which yields a virus that is severely attenuated in cell culture.
RESUMO
In autumn 2011, a novel species of the genus Orthobunyavirus of the Simbu serogroup was discovered close to the German/Dutch border and named Schmallenberg virus (SBV). Since then, SBV has caused a large epidemic in European livestock. Like other viruses of the Simbu serogroup, SBV is transmitted by insect vectors. Adult ruminants may show a mild transient disease, while an infection during a critical period of pregnancy can lead to severe congenital malformation, premature birth or stillbirth. The current knowledge about the virus, its diagnosis, the spread of the epidemic, the impact and the possibilities for preventing infections with SBV is described and discussed.
Assuntos
Doenças dos Animais/epidemiologia , Doenças dos Animais/virologia , Infecções por Bunyaviridae/veterinária , Orthobunyavirus , Doenças dos Animais/diagnóstico , Doenças dos Animais/prevenção & controle , Doenças dos Animais/transmissão , Animais , Infecções por Bunyaviridae/diagnóstico , Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/prevenção & controle , Infecções por Bunyaviridae/transmissão , Ceratopogonidae/virologia , Anormalidades Congênitas/veterinária , Anormalidades Congênitas/virologia , Europa (Continente)/epidemiologia , Insetos Vetores/virologia , Orthobunyavirus/genética , Orthobunyavirus/patogenicidade , Reação em Cadeia da Polimerase em Tempo Real , Ruminantes/virologia , Estudos SoroepidemiológicosRESUMO
Schmallenberg virus (SBV), a novel orthobunyavirus, was discovered in Germany in 2011. In adult ruminants SBV causes mild transient disease, but foetal infection can lead to severe malformations. Owing to its recent discovery, the knowledge about the pathogenesis is limited. In this study, two heifers seroconverted after a previous SBV infection and five SBV antibody-negative calves were subcutaneously inoculated, another two animals received SBV orally and three were kept as controls. In naïve cattle infected subcutaneously viral RNA was detected in serum and blood samples for several days. Seropositive or orally inoculated animals as well as the uninfected controls remained negative throughout the study. Seroconversion was observed only after subcutaneous exposure of the naïve animals to SBV. In lymphocytes from peripheral blood SBV genome was not detected, but the lymphocyte homeostasis in blood was influenced.