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2.
Arch Virol ; 166(3): 881-884, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33433694

RESUMO

In the present study, we serosurveyed the exposure of 222 draft horses to different arboviruses in the city of Santa Fe, Argentina. Plaque reduction neutralization tests confirmed exposure to Fort Sherman virus (FSV), Saint Louis encephalitis virus (SLEV), West Nile virus (WNV), and Río Negro virus (RNV). Apparently, Western and Eastern equine encephalitis viruses did not circulate in the population tested. The confirmation of five seroconversions for WNV, FSV, and SLEV and the association between prevalence and age are indicative of recent circulation. These results highlight the importance of considering draft horses in arboviral surveillance in urban and rural areas of developing countries.


Assuntos
Infecções por Alphavirus/epidemiologia , Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/epidemiologia , Encefalite de St. Louis/epidemiologia , Doenças dos Cavalos/epidemiologia , Febre do Nilo Ocidental/epidemiologia , Alphavirus/imunologia , Alphavirus/isolamento & purificação , Infecções por Alphavirus/veterinária , Animais , Argentina/epidemiologia , Infecções por Bunyaviridae/veterinária , Vírus da Encefalite de St. Louis/imunologia , Vírus da Encefalite de St. Louis/isolamento & purificação , Encefalite de St. Louis/veterinária , Doenças dos Cavalos/virologia , Cavalos , Orthobunyavirus/imunologia , Orthobunyavirus/isolamento & purificação , Soroconversão , Febre do Nilo Ocidental/veterinária , Vírus do Nilo Ocidental/imunologia , Vírus do Nilo Ocidental/isolamento & purificação
3.
J Virol ; 94(17)2020 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-32522852

RESUMO

Schmallenberg virus (SBV) is an insect-transmitted orthobunyavirus that can cause abortions and congenital malformations in the offspring of ruminants. Even though the two viral surface glycoproteins Gn and Gc are involved in host cell entry, the specific cellular receptors of SBV are currently unknown. Using genome-wide CRISPR-Cas9 forward screening, we identified 3'-phosphoadenosine 5'-phosphosulfate (PAPS) transporter 1 (PAPST1) as an essential factor for SBV infection. PAPST1 is a sulfotransferase involved in heparan sulfate proteoglycan synthesis encoded by the solute carrier family 35 member B2 gene (SLC35B2). SBV cell surface attachment and entry were largely reduced upon the knockout of SLC35B2, whereas the reconstitution of SLC35B2 in these cells fully restored their susceptibility to SBV infection. Furthermore, treatment of cells with heparinase diminished infection with SBV, confirming that heparan sulfate plays an important role in cell attachment and entry, although to various degrees, heparan sulfate was also found to be important to initiate infection by two other bunyaviruses, La Crosse virus and Rift Valley fever virus. Thus, PAPST1-triggered synthesis of cell surface heparan sulfate is required for the efficient replication of SBV and other bunyaviruses.IMPORTANCE SBV is a newly emerging orthobunyavirus (family Peribunyaviridae) that has spread rapidly across Europe since 2011, resulting in substantial economic losses in livestock farming. In this study, we performed unbiased genome-wide CRISPR-Cas9 screening and identified PAPST1, a sulfotransferase encoded by SLC35B2, as a host entry factor for SBV. Consistent with its role in the synthesis of heparan sulfate, we show that this activity is required for efficient infection by SBV. A comparable dependency on heparan sulfate was also observed for La Crosse virus and Rift Valley fever virus, highlighting the importance of heparan sulfate for host cell infection by bunyaviruses. Thus, the present work provides crucial insights into virus-host interactions of important animal and human pathogens.


Assuntos
Infecções por Bunyaviridae/genética , Infecções por Bunyaviridae/virologia , Sistemas CRISPR-Cas , Orthobunyavirus/genética , Orthobunyavirus/fisiologia , Animais , Bunyaviridae , Chlorocebus aethiops , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Europa (Continente) , Técnicas de Inativação de Genes , Células HEK293 , Heparitina Sulfato/metabolismo , Humanos , Gado , Glicoproteínas de Membrana/genética , Orthobunyavirus/patogenicidade , Vírus da Febre do Vale do Rift , Transportadores de Sulfato/metabolismo , Sulfotransferases/metabolismo , Células Vero , Ligação Viral
4.
PLoS One ; 15(6): e0233722, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32479526

RESUMO

Melon yellow spot orthotospovirus (MYSV), a member of the genus Orthotospovirus, is an important virus in cucurbits. Thrips palmi is considered the most serious pest of cucurbits because it directly damages and indirectly transmits MYSV to the plant. The effects of MYSV-infected plants on the development time, fecundity, and preference of the thrips were analyzed in this study. Our results showed that the development time of male and female thrips did not differ significantly between MYSV-infected and non-infected cucumbers. The survival rate of thrips in non-infected and MYSV-infected cucumbers were not significantly different. In a non-choice assay, T. palmi adults were released on non-infected and MYSV-infected cucumbers and allowed to lay eggs. The number of hatched larvae did not significantly differ between non-infected and MYSV-infected cucumbers. In a choice assay, MYSV had no detectable effect on the number of adult thrips and preceding hatched larvae. In a pull assay, the settling rate of thrips on the released plant did not differ significantly when the adult thrips were released to non-infected or MYSV infected cucumbers for any cucumber cultivar. Based on our results, we propose that the effects of MYSV-infected cucumbers on the development time, fecundity, or preference of T. palmi may not be an important factor in MYSV spread between cucumbers.


Assuntos
Cucumis sativus/parasitologia , Orthobunyavirus/patogenicidade , Tisanópteros/fisiologia , Animais , Feminino , Fertilidade , Especificidade de Hospedeiro , Masculino , Tisanópteros/crescimento & desenvolvimento , Tisanópteros/patogenicidade , Tisanópteros/virologia
5.
Am J Trop Med Hyg ; 103(1): 183-189, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32314686

RESUMO

In 2018, a large outbreak of Rift Valley fever (RVF)-like illness in cattle in Rwanda and surrounding countries was reported. From this outbreak, sera samples from 157 cows and 28 goats suspected to be cases of RVF were tested to confirm or determine the etiology of the disease. Specifically, the hypothesis that orthobunyaviruses-Bunyamwera virus (BUNV), Batai virus (BATV), and Ngari virus (NRIV)-were co-circulating and contributed to RVF-like disease was tested. Using reverse transcriptase-polymerase chain reaction (RT-PCR), RVFV RNA was detected in approximately 30% of acutely ill animals, but in all cases of hemorrhagic disease. Seven cows with experienced abortion had positive amplification and visualization by gel electrophoresis of all three segments of either BUNV or BATV, and three of these were suggested to be coinfected with BUNV and BATV. On sequencing, five of these seven cows were conclusively positive for BUNV. However, in several other animals, sequencing was successful for some but not all segments of targeted viruses BUNV and BATV. In addition, there was evidence of RVFV-orthobunyavirus coinfection, through RT-PCR/gel electrophoresis and subsequent Sanger sequencing. In no cases were we able to definitely identify the specific coinfecting viral species. This is the first time evidence for orthobunyavirus circulation has been molecularly confirmed in Rwanda. Furthermore, RT-PCR results suggest that BUNV and BATV may coinfect cattle and that RVFV-infected animals may be coinfected with other orthobunyaviruses. Finally, we confirm that BUNV and, perhaps, other orthobunyaviruses were co-circulating with RVFV and contributed to the burden of disease attributed to RVFV in Rwanda.


Assuntos
Vírus Bunyamwera/genética , Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/epidemiologia , Surtos de Doenças , Orthobunyavirus/genética , Febre do Vale de Rift/epidemiologia , Vírus da Febre do Vale do Rift/genética , Animais , Vírus Bunyamwera/classificação , Vírus Bunyamwera/isolamento & purificação , Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/transmissão , Infecções por Bunyaviridae/virologia , Bovinos , Doenças dos Bovinos/transmissão , Doenças dos Bovinos/virologia , Coinfecção , Feminino , Cabras/virologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Epidemiologia Molecular , Orthobunyavirus/classificação , Orthobunyavirus/isolamento & purificação , RNA Viral/genética , Febre do Vale de Rift/transmissão , Febre do Vale de Rift/virologia , Vírus da Febre do Vale do Rift/classificação , Vírus da Febre do Vale do Rift/isolamento & purificação , Ruanda/epidemiologia
6.
Artigo em Inglês | MEDLINE | ID: mdl-32284379

RESUMO

Bunyaviruses are significant human pathogens, causing diseases ranging from hemorrhagic fevers to encephalitis. Among these viruses, La Crosse virus (LACV), a member of the California serogroup, circulates in the eastern and midwestern United States. While LACV infection is often asymptomatic, dozens of cases of encephalitis are reported yearly. Unfortunately, no antivirals have been approved to treat LACV infection. Here, we developed a method to rapidly test potential antivirals against LACV infection. From this screen, we identified several potential antiviral molecules, including known antivirals. Additionally, we identified many novel antivirals that exhibited antiviral activity without affecting cellular viability. Valinomycin, a potassium ionophore, was among our top targets. We found that valinomycin exhibited potent anti-LACV activity in multiple cell types in a dose-dependent manner. Valinomycin did not affect particle stability or infectivity, suggesting that it may preclude virus replication by altering cellular potassium ions, a known determinant of LACV entry. We extended these results to other ionophores and found that the antiviral activity of valinomycin extended to other viral families, including bunyaviruses (Rift Valley fever virus, Keystone virus), enteroviruses (coxsackievirus, rhinovirus), flavirivuses (Zika virus), and coronaviruses (human coronavirus 229E [HCoV-229E] and Middle East respiratory syndrome CoV [MERS-CoV]). In all viral infections, we observed significant reductions in virus titer in valinomycin-treated cells. In sum, we demonstrate the importance of potassium ions to virus infection, suggesting a potential therapeutic target to disrupt virus replication.


Assuntos
Antivirais/farmacologia , Encefalite da Califórnia/tratamento farmacológico , Ionóforos/farmacologia , Vírus La Crosse/efeitos dos fármacos , Potássio/metabolismo , Valinomicina/farmacologia , Replicação Viral/efeitos dos fármacos , Coronavirus/efeitos dos fármacos , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Encefalite da Califórnia/virologia , Enterovirus/efeitos dos fármacos , Flavivirus/efeitos dos fármacos , Humanos , Orthobunyavirus/efeitos dos fármacos , Estados Unidos
8.
Mem Inst Oswaldo Cruz ; 115: e190338, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32130368

RESUMO

Oropouche virus (OROV) is an arthropod-borne virus of the Peribunyaviridae family, transmitted to humans primarily by Culicoides paraensis. It is one of the main arboviruses infecting humans in Brazil, primarily in the Amazon Region. Here, we report the detection of OROV in the saliva and urine of a patient whose samples were collected five days after the onset of symptoms. Nucleotide sequencing and phylogenetic analysis further confirmed the results. To our knowledge, this is the first study reporting the detection of OROV in the saliva and urine of an infected patient. In addition, the results of our study expand the current knowledge pertaining to the natural history of Oropouche fever.


Assuntos
Infecções por Bunyaviridae/diagnóstico , Orthobunyavirus/genética , Orthobunyavirus/isolamento & purificação , Saliva/virologia , Urina/virologia , Sequência de Aminoácidos , Sequência de Bases , Feminino , Humanos , Pessoa de Meia-Idade , Filogenia , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa
9.
Virus Genes ; 56(2): 150-167, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32076918

RESUMO

The viruses historically implicated or currently considered as candidates for misuse in bioterrorist events are poxviruses, filoviruses, bunyaviruses, orthomyxoviruses, paramyxoviruses and a number of arboviruses causing encephalitis, including alpha- and flaviviruses. All these viruses are of concern for public health services when they occur in natural outbreaks or emerge in unvaccinated populations. Recent events and intelligence reports point to a growing risk of dangerous biological agents being used for nefarious purposes. Public health responses effective in natural outbreaks of infectious disease may not be sufficient to deal with the severe consequences of a deliberate release of such agents. One important aspect of countermeasures against viral biothreat agents are the antiviral treatment options available for use in post-exposure prophylaxis. These issues were adressed by the organizers of the 16th Medical Biodefense Conference, held in Munich in 2018, in a special session on the development of drugs to treat infections with viruses currently perceived as a threat to societies or associated with a potential for misuse as biothreat agents. This review will outline the state-of-the-art methods in antivirals research discussed and provide an overview of antiviral compounds in the pipeline that are already approved for use or still under development.


Assuntos
Antivirais/uso terapêutico , Arbovirus/efeitos dos fármacos , Bioterrorismo/prevenção & controle , Viroses/tratamento farmacológico , Arbovirus/patogenicidade , Filoviridae/efeitos dos fármacos , Filoviridae/patogenicidade , Humanos , Orthobunyavirus/efeitos dos fármacos , Orthobunyavirus/patogenicidade , Orthomyxoviridae/efeitos dos fármacos , Orthomyxoviridae/patogenicidade , Paramyxovirinae/efeitos dos fármacos , Paramyxovirinae/patogenicidade , Poxviridae/efeitos dos fármacos , Poxviridae/patogenicidade , Viroses/virologia
10.
BMC Res Notes ; 13(1): 67, 2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-32041646

RESUMO

OBJECTIVE: To evaluate the frequency of infection caused by the Oropouche virus (OROV) in 496 patients with acute febrile disease (AFI), whose samples were obtained for the analysis of endemic arboviruses in a previous investigation carried out in 2016. RESULTS: OROV was detected in 26.4% (131/496) of serum samples from patients with AFI. Co-infections with Dengue virus (7.3%), Zika virus (1.8%) and Chikungunya (0.2%) were observed. The most common clinical symptoms reported among the patients with OROV infections were headache 85.5% (112/131), myalgia 80.9% (106/131), arthralgia 72.5% (95/131) and loss of appetite 67.9% (89/131). Headache and myalgia were predominant in all age groups. Both OROV infections and co-infections were more frequent in May, June and July corresponding to the dry season of the region.


Assuntos
Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/fisiopatologia , Febre de Chikungunya/epidemiologia , Coinfecção , Dengue/epidemiologia , Orthobunyavirus/patogenicidade , Infecção por Zika virus/epidemiologia , Adolescente , Adulto , Idoso , Infecções por Bunyaviridae/sangue , Infecções por Bunyaviridae/complicações , Febre de Chikungunya/sangue , Criança , Pré-Escolar , Comorbidade , Dengue/sangue , Feminino , Febre/epidemiologia , Febre/etiologia , Humanos , Masculino , Pessoa de Meia-Idade , Peru/epidemiologia , Adulto Jovem , Infecção por Zika virus/sangue
11.
Transbound Emerg Dis ; 67(4): 1433-1441, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32009301

RESUMO

An orthobunyavirus termed Fort Sherman virus (FSV) was isolated in 1985 from a febrile US soldier in Panama, yet potential animal reservoirs remained unknown. We investigated sera from 192 clinically healthy peri-domestic animals sampled in northeastern Brazil during 2014-2018 by broadly reactive RT-PCR for orthobunyavirus RNA, including 50 cattle, 57 sheep, 35 goats and 50 horses. One horse sampled in 2018 was positive (0.5%; 95% CI, 0.01-3.2) at 6.2 × 103 viral RNA copies/mL. Genomic comparisons following virus isolation in Vero cells and deep sequencing revealed high identity of translated amino acid sequences between the new orthobunyavirus and the Panamanian FSV prototype (genes: L, 98.8%; M, 83.5%; S, 100%), suggesting these viruses are conspecific. Database comparisons revealed even higher genomic identity between the Brazilian FSV and taxonomically unassigned Argentinian mosquito- and horse-derived viruses sampled in 1965, 1982 and 2013 with only 1.1% maximum translated amino acid distances across viral genes, suggesting the Argentinian viruses were also distinct FSV strains. The Panamanian FSV strain was an M gene reassortant relative to all Southern American FSV strains, clustering phylogenetically with Cache Valley virus (CVV). Mean dN/dS ratios among FSV genes ranged from 0.03 to 0.07, compatible with strong purifying selection. FSV-specific neutralizing antibodies occurred at relatively high end-point titres in the range of 1:300 in 22.0% of horses (11 out of 50 animals), 8.0% of cattle (4/50 animals), 7.0% of sheep (4/57 animals) and 2.9% of goats (1/35 animals). High specificity of serologic testing was suggested by significantly higher overall FSV-specific compared to CVV- and Bunyamwera virus-specific end-point titres (p = .009), corroborating a broad vertebrate host range within peri-domestic animals. Growth kinetics using mosquito-, midge- and sandfly-derived cell lines suggested Aedes mosquitos as potential vectors. Our findings highlight the occurrence of FSV across a geographic range exceeding 7,000 km, surprising genomic conservation across a time span exceeding 50 years, M gene-based reassortment events, and the existence of multiple animal hosts of FSV.


Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/virologia , Doenças das Cabras/virologia , Doenças dos Cavalos/virologia , Mosquitos Vetores/virologia , Orthobunyavirus/isolamento & purificação , Doenças dos Ovinos/virologia , Aedes/virologia , Animais , Brasil , Infecções por Bunyaviridae/virologia , Bovinos , Chlorocebus aethiops , Cabras , Cavalos , Especificidade de Hospedeiro , Orthobunyavirus/genética , Filogenia , Ovinos , Células Vero , Zoonoses
12.
PLoS Negl Trop Dis ; 14(1): e0007897, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31961856

RESUMO

Oropouche virus (OROV) is responsible for outbreaks of Oropouche fever in parts of South America. We recently identified and isolated OROV from a febrile Ecuadorian patient, however, a previously published qRT-PCR assay did not detect OROV in the patient sample. A primer mismatch to the Ecuadorian OROV lineage was identified from metagenomic sequencing data. We report the optimisation of an qRT-PCR assay for the Ecuadorian OROV lineage, which subsequently identified a further five cases in a cohort of 196 febrile patients. We isolated OROV via cell culture and developed an algorithmically-designed primer set for whole-genome amplification of the virus. Metagenomic sequencing of the patient samples provided OROV genome coverage ranging from 68-99%. The additional cases formed a single phylogenetic cluster together with the initial case. OROV should be considered as a differential diagnosis for Ecuadorian patients with febrile illness to avoid mis-diagnosis with other circulating pathogens.


Assuntos
Infecções por Bunyaviridae/virologia , Orthobunyavirus/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Infecções por Bunyaviridae/diagnóstico , Estudos de Coortes , Equador , Genoma Viral , Humanos , Metagenoma , Orthobunyavirus/classificação , Orthobunyavirus/genética , Filogenia , RNA Viral/genética
13.
Transbound Emerg Dis ; 67(4): 1708-1715, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31991522

RESUMO

Schmallenberg virus (SBV) is a vector-borne virus belonging to the genus Orthobunyavirus within the Bunyaviridae family. SBV emerged in Europe in 2011 and was characterized by epidemics of abortions, stillbirths and congenital malformations in domestic ruminants. The first evidence of SBV infection in Slovenia was from an ELISA-positive sample from a cow collected in August 2012; clinical manifestations of SBV disease in sheep and cattle were observed in 2013, with SBV RNA detected in samples collected from a total of 28 herds. A potential re-emergence of SBV in Europe is predicted to occur when population-level immunity declines. SBV is also capable of infecting several wild ruminant species, although clinical disease has not yet been described in these species. Data on SBV-positive wild ruminants suggest that these species might be possible sources for the re-emergence of SBV. The aim of this study was to investigate whether SBV was circulating among wild ruminants in Slovenia and whether these species can act as a virus reservoir. A total of 281 blood and spleen samples from wild ruminants, including roe deer, red deer, chamois and European mouflon, were collected during the 2017-2018 hunting season. Serum samples were tested for antibodies against SBV by ELISA; the overall seroprevalence was 18.1%. Seropositive samples were reported from all over the country in examined animal species from 1 to 15 years of age. Spleen samples from the seropositive animals and serum samples from the seronegative animals were tested for the presence of SBV RNA using real-time RT-PCR; all the samples tested negative. Based on the results of the seropositive animals, it was demonstrated that SBV was circulating in wild ruminant populations in Slovenia even after the epidemic, as almost half (23/51) of the seropositive animals were 1 or 2 years old.


Assuntos
Animais Selvagens/virologia , Infecções por Bunyaviridae/veterinária , Orthobunyavirus/isolamento & purificação , Ruminantes/virologia , Animais , Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/virologia , Cervos/virologia , Reservatórios de Doenças/veterinária , Reservatórios de Doenças/virologia , Ensaio de Imunoadsorção Enzimática/veterinária , Epidemias/veterinária , Orthobunyavirus/genética , Orthobunyavirus/imunologia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Rupicapra/virologia , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/virologia , Carneiro Doméstico/virologia , Eslovênia/epidemiologia
14.
Jpn J Infect Dis ; 73(2): 164-165, 2020 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-31787741

RESUMO

Oropouche virus (OROV) is a negative-sense, single-stranded RNA arbovirus transmitted to humans by the midge Culicoides paraenesis, causing Oropouche fever. Reports of its outbreak in Brazil have so far been restricted to the Central-Northern region of the country. However, its incidence is underestimated, mainly due to its clinical similarities with other arbovirus diseases, including dengue (DENV), chikungunya (CHIKV), and zika (ZIKV), and the lack of specific diagnostic tests. Here, we report for the first time, the detection of OROV in saliva and urine samples, and cases of autochthone OROV infections in Salvador Metropolitan region, Bahia, a Northeastern capital in the coast of Brazil. Serum, saliva, and urine samples negative for DENV, CHIKV, and ZIKV were tested for OROV using a reverse transcription nested polymerase chain reaction (RT-nested-PCR) protocol, and 2 serum, 2 saliva, and 1 urine samples were positive. This report shows the need for an efficient surveillance system for controlling the spread of this virus, and suggests the use of saliva and urine as alternative samples for OROV detection in the absence of serum samples.


Assuntos
Infecções por Bunyaviridae/diagnóstico , Infecções por Bunyaviridae/urina , Febre/virologia , Orthobunyavirus/genética , Saliva/virologia , Animais , Brasil/epidemiologia , Infecções por Bunyaviridae/epidemiologia , Ceratopogonidae/virologia , Surtos de Doenças , Humanos , Orthobunyavirus/isolamento & purificação , RNA Viral/genética
15.
J Virol ; 94(5)2020 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-31801869

RESUMO

The Amazon basin is home to numerous arthropod-borne viral pathogens that cause febrile disease in humans. Among these, Oropouche orthobunyavirus (OROV) is a relatively understudied member of the genus Orthobunyavirus, family Peribunyaviridae, that causes periodic outbreaks in human populations in Brazil and other South American countries. Although several studies have described the genetic diversity of the virus, the evolutionary processes that shape the OROV genome remain poorly understood. Here, we present a comprehensive study of the genomic dynamics of OROV that encompasses phylogenetic analysis, evolutionary rate estimates, inference of natural selective pressures, recombination and reassortment, and structural analysis of OROV variants. Our study includes all available published sequences, as well as a set of new OROV genome sequences obtained from patients in Ecuador, representing the first set of genomes from this country. Our results show differing evolutionary processes on the three segments that comprise the viral genome. We infer differing times of the most recent common ancestors of the genome segments and propose that this can be explained by cryptic reassortment. We also present the discovery of previously unobserved putative N-linked glycosylation sites, as well as codons that evolve under positive selection on the viral surface proteins, and discuss the potential role of these features in the evolution of OROV through a combined phylogenetic and structural approach.IMPORTANCE The emergence and reemergence of pathogens such as Zika virus, chikungunya virus, and yellow fever virus have drawn attention toward other cocirculating arboviruses in South America. Oropouche virus (OROV) is a poorly studied pathogen responsible for over a dozen outbreaks since the early 1960s and represents a public health burden to countries such as Brazil, Panama, and Peru. OROV is likely underreported since its symptomatology can be easily confounded with other febrile illnesses (e.g., dengue fever and leptospirosis) and point-of-care testing for the virus is still uncommon. With limited data, there is a need to optimize the information currently available. Analysis of OROV genomes can help us understand how the virus circulates in nature and can reveal the evolutionary forces that shape the genetic diversity of the virus, which has implications for molecular diagnostics and the design of potential vaccines.


Assuntos
Evolução Molecular , Genoma Viral , Orthobunyavirus/classificação , Orthobunyavirus/genética , Filogenia , Infecções por Bunyaviridae/epidemiologia , Infecções por Bunyaviridae/virologia , Equador , Humanos , Modelos Moleculares , Conformação Proteica , Seleção Genética , América do Sul , Proteínas Virais/química , Proteínas Virais/genética , Sequenciamento Completo do Genoma
16.
Diagn Microbiol Infect Dis ; 96(1): 114894, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31727377

RESUMO

Oropouche virus (OROV) causes an acute, systemic febrile illness, and in certain regions of South America, this represents the second most common human arboviral infection after dengue virus. A new real-time RT-PCR was developed for OROV and reassortant species. The new OROV rRT-PCR proved linear across 6-7 orders of magnitude with a lower limit of 95% detection of 5.6-10.8 copies/µL. Upon testing dilutions of OROV and Iquitos virus reference genomic RNA, all dilutions with >10 copies/µL were detected in both the OROV rRT-PCR and a comparator molecular assay, but the OROV rRT-PCR detected more samples with ≤10 copies/µL (8/14 vs 0/13, respectively, P = 0.002). In a set of 100 acute-phase clinical samples from Paraguay patients with a suspected arboviral illness, no patients tested positive for OROV RNA using either assay. The OROV rRT-PCR provides a sensitive molecular assay for the study of this important yet neglected tropical arboviral infection.


Assuntos
Infecções por Bunyaviridae/diagnóstico , Orthobunyavirus/isolamento & purificação , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Adulto , Infecções por Bunyaviridae/virologia , Feminino , Humanos , Limite de Detecção , Masculino , Pessoa de Meia-Idade , Paraguai , Sensibilidade e Especificidade
17.
Artigo em Inglês | MEDLINE | ID: mdl-31835858

RESUMO

The outbreak of a previously unknown and new disease in the United Kingdom, known as 'Schmallenberg disease', a disease associated with abortions, stillbirths and fetal deformities in naïve ewes, was reported for the first time in South West England during the 2012/13 early lambing season. Epidemiological studies confirmed that the Schmallenberg virus (SBV) had a severe negative impact upon animal welfare and the productivity of affected flocks. By contrast, there was a specific lack of research on the impact of SBV on sheep farmer well-being. This study aimed to improve our understanding of sheep farmers' experiences of Schmallenberg disease, and the impact of the first outbreak on sheep farmer well-being during the 2012/13 early lambing season in South West England. Face-to-face, semi-structured interviews with six farmers with small flocks of pedigree and purebred sheep in South West England were conducted in 2013. The data were analysed via thematic analysis. The main themes regarding the impact of disease on farmer well-being included: (i) emotional highs and lows are part of a normal lambing season; (ii) negative emotions and memories associated with the Schmallenberg disease outbreak; and (iii) resilience and coping with the unexpected disease outbreak. These novel data present preliminary findings from a small number of sheep farmers, and indicate that for some farmers, an unexpected outbreak of a new and emerging disease for the first time during lambing, and dealing with high levels of dystocia, deformities and deaths in their animals, had a negative impact on their emotional well-being during the peak period of the sheep production cycle.


Assuntos
Infecções por Bunyaviridae/veterinária , Surtos de Doenças/veterinária , Fazendeiros/psicologia , Doenças dos Ovinos , Adaptação Psicológica , Animais , Emoções , Inglaterra , Feminino , Humanos , Masculino , Orthobunyavirus , Gravidez , Ovinos , Natimorto/veterinária
18.
J Vet Sci ; 20(6): e58, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31775185

RESUMO

The Schmallenberg virus (SBV) is an orthobunyavirus that causes abortions, stillbirths, and congenital defects in pregnant sheep and cattle. Inactivated or live attenuated vaccines have been developed in endemic countries, but there is still interest in the development of SBV vaccines that would allow Differentiating Infected from Vaccinated Animals (DIVA). Therefore, an attempt was made to develop novel DIVA-compatible SBV vaccines using SBV glycoproteins expressed in baculovirus. All vaccines and phosphate buffered saline (PBS) controls were prepared with adjuvant and administered subcutaneously to cattle at 6 month of age. The first trial included 2 groups of animals vaccinated with either carboxyl-terminus glycoprotein (Gc) or PBS and boosted after 2 weeks. In the second trial, 3 groups of cattle were administered either Gc, Gc and amino-terminus glycoprotein (Gn), or PBS with a booster vaccination after 3 weeks. The animals were challenged with SBV 9 days after the booster vaccination in the first study, and 3 weeks after the booster vaccination in the second study. Using a SBV Gc-specific enzyme-linked immunosorbent assay, antibodies were first detected in serum samples 14 days after the first vaccination in both trials, and peaked on days 7 and 9 after the booster in the first and second trials, respectively. Low titers of neutralizing antibodies were detected in serum from only 3/6 and 2/4 animals in the first and second trial, respectively, at 14 days after the first vaccination. The titers increased 2 to 3-fold after the booster vaccination. SBV-specific RNA was detected in the serum and selective tissues in all animals after SBV challenge independent of vaccination status. The SBV candidate vaccines neither prevented viremia nor conferred protection against SBV infection.


Assuntos
Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/prevenção & controle , Glicoproteínas/imunologia , Imunogenicidade da Vacina , Orthobunyavirus/fisiologia , Proteínas do Envelope Viral/imunologia , Vacinas Virais/imunologia , Animais , Infecções por Bunyaviridae/prevenção & controle , Bovinos , Doenças dos Bovinos/imunologia , Distribuição Aleatória , Vacinação/veterinária , Vacinas de Subunidades/imunologia
19.
BMC Vet Res ; 15(1): 426, 2019 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-31779623

RESUMO

BACKGROUND: Schmallenberg virus (SBV) is a midge borne virus of cattle and sheep. Infection is typically asymptomatic in adult sheep but fetal infection during pregnancy can result in abortion, stillbirth, neurological disorders and malformations of variable severity in newborn animals. It was first identified in Germany and the Netherlands in 2011 and then circulated throughout Europe in 2012 and 2013. Circulation in subsequent years was low or non-existent until summer and autumn 2016, leading to an increased incidence of deformed newborn lambs and calves in 2016-17. This study reports SBV circulation in October 2016 within a group of 24 ewes and 13 rams. The ewes were monitored at 3 times points over an 11 week period (September to December 2016). RESULTS: Most ewes displayed an increase in SBV VNT with antibody titre increases greater in older, previously exposed ewes. Two ewes had SBV RNA detectable by RT-qPCR, one on 30/09/16 and one on 04/11/16. Of these ewes, one had detectable serum SBV RNA (indicating viraemia) despite pre-existing antibody. The rams had been previously vaccinated with a commercial inactivated SBV vaccine, they showed minimal neutralising antibody titres against SBV 8 months post-vaccination and all displayed increased titre in October 2016. CONCLUSION: This data suggests that SBV circulated for a minimum period of 5 weeks in September to October 2016 in central England. Ewes previously exposed to virus showed an enhanced antibody response compared to naïve animals. Pre-existing antibody titre did not prevent re-infection in at least one animal, implying immunity to SBV upon natural exposure may not be life-long. In addition, data suggests that immunity provided by killed adjuvanted SBV vaccines only provides short term protection (< 8 months) from virus.


Assuntos
Infecções por Bunyaviridae/veterinária , Orthobunyavirus/imunologia , Doenças dos Ovinos/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/sangue , Infecções por Bunyaviridae/imunologia , Inglaterra/epidemiologia , Feminino , Masculino , RNA Viral/sangue , Ovinos , Doenças dos Ovinos/virologia , Carneiro Doméstico , Vacinação
20.
BMC Vet Res ; 15(1): 408, 2019 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-31711494

RESUMO

BACKGROUND: Akabane disease (AD), a barrier to international trade for endemic areas with far economic impact on the countries, is caused by Akabane virus (AKAV). Commercial enzyme-linked immunosorbent assay (ELISA) is a commonly used diagnostic technique for AKAV infection, including the IDEXX and IDVET ELISA kits. However, the comparative evaluation of the IDEXX and IDVET ELISA kits has not been published. The object of this study was to evaluate the test performance of the two commercial ELISA kits in detecting serum anti-AKAV antibodies in cattle. RESULTS: With virus neutralization test (VNT) as the "relative gold standard", the diagnostic sensitivity (DSe) was 80.39% (123/153) and 93.46% (143/153) for the IDEXX and IDVET ELISA kit, when suspect samples were included. The diagnostic specificity (DSp) for the IDEXX and IDVET ELISA kit was 93.48% (502/537) and 82.31% (442/537), respectively. CONCLUSION: Both of the tested ELISA kits could be applied to detect antibodies against AKAV in cattle serum. The IDVET ELISA kit had a higher DSe. The IDEXX ELISA kit possessed the higher DSp. These results have important implications if the kits are used to screen herds or individual cattle in surveillance programs, or at border crossings for import-export inspection and quarantine.


Assuntos
Anticorpos Antivirais/sangue , Infecções por Bunyaviridae/veterinária , Doenças dos Bovinos/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Orthobunyavirus/imunologia , Animais , Infecções por Bunyaviridae/diagnóstico , Infecções por Bunyaviridae/imunologia , Bovinos , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/virologia , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Testes de Neutralização/veterinária , Sensibilidade e Especificidade
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