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1.
Emerg Infect Dis ; 19(2): 264-6, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23347828

RESUMO

We investigated viability of hepatitis E virus (HEV) identified in contaminated pork liver sausages obtained from France. HEV replication was demonstrated in 1 of 4 samples by using a 3-dimensional cell culture system. The risk for human infection with HEV by consumption of these sausages should be considered to be high.


Assuntos
Vírus da Hepatite E/fisiologia , Produtos da Carne/virologia , Vírion/fisiologia , Animais , Linhagem Celular Tumoral , Microbiologia de Alimentos , França , Vírus da Hepatite E/genética , Vírus da Hepatite E/ultraestrutura , Humanos , Fígado , Tipagem Molecular , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sus scrofa , Vírion/genética , Vírion/ultraestrutura , Replicação Viral
2.
Emerg Infect Dis ; 18(8): 1358-60, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22840183

RESUMO

We investigated contamination by hepatitis E virus (HEV) in the pork production chain in the United Kingdom. We detected HEV in pig liver samples in a slaughterhouse, in surface samples from a processing plant, and in pork sausages and surface samples at point of sale. Our findings provide evidence for possible foodborne transmission of HEV during pork production.


Assuntos
Manipulação de Alimentos/métodos , Microbiologia de Alimentos , Vírus da Hepatite E/isolamento & purificação , Hepatite E/transmissão , Suínos/virologia , Matadouros , Animais , Fezes/virologia , Hepatite E/virologia , Vírus da Hepatite E/genética , Fígado/virologia , Produtos da Carne/virologia , RNA Viral/genética , Doenças dos Suínos/transmissão , Doenças dos Suínos/virologia , Reino Unido/epidemiologia
3.
Lancet Infect Dis ; 8(11): 698-709, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18992406

RESUMO

Hepatitis E is endemic in many developing countries where it causes substantial morbidity. In industrialised countries, it is considered rare, and largely confined to travellers returning from endemic areas. However, there is now a growing body of evidence that challenges this notion. Autochthonous hepatitis E in developed countries is far more common than previously recognised, and might be more common than hepatitis A. Hepatitis E has a predilection for older men in whom it causes substantial morbidity and mortality. The disease has a poor prognosis in the context of pre-existing chronic liver disease, and is frequently misdiagnosed as drug-induced liver injury. The source and route of infection remain uncertain, but it might be a porcine zoonosis. Patients with unexplained hepatitis should be tested for hepatitis E, whatever their age or travel history.


Assuntos
Doenças Transmissíveis Emergentes/epidemiologia , Países Desenvolvidos , Hepatite E/epidemiologia , Animais , Doenças Transmissíveis Emergentes/diagnóstico , Doenças Transmissíveis Emergentes/terapia , Hepatite E/diagnóstico , Hepatite E/terapia , Vírus da Hepatite E/classificação , Humanos , Incidência , Estudos Soroepidemiológicos , Zoonoses/transmissão
4.
Eur J Gastroenterol Hepatol ; 20(8): 800-3, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18617787

RESUMO

Hepatitis E virus is endemic in many parts of the developing world and causes a self-limiting hepatitis in young adults, except in pregnant women and patients with chronic liver disease, where the mortality is high. Locally acquired hepatitis E is increasingly recognized in the developed world. It is caused by hepatitis E virus genotype 3, affects the middle-aged and the elderly, and may be a zoonotic infection from pigs. We present a case of locally acquired hepatitis E infection in a patient with previously undiagnosed cirrhosis, which resulted in subacute liver failure and death. We describe our attempt to trace this infection to a free-range pig farm adjacent to the patient's place of employment. Hepatitis E infection should be considered in the differential diagnosis in patients with decompensated chronic liver disease whatever their age or travel history. When found, the prognosis may be poor.


Assuntos
Hepatite E/diagnóstico , Cirrose Hepática/complicações , Animais , Doença Crônica , Evolução Fatal , Hepatite E/complicações , Hepatite E/transmissão , Hepatite E/virologia , Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Humanos , Masculino , Pessoa de Meia-Idade , Sus scrofa , Zoonoses/transmissão
5.
Eur J Gastroenterol Hepatol ; 20(8): 784-90, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18617784

RESUMO

AIMS: To report the natural history of autochthonous hepatitis E and hepatitis E virus (HEV) IgG seroprevalence in Southwest England. METHODS: Patients with unexplained hepatitis were tested for hepatitis E and cases followed until recovery or death. Five hundred blood donors, 336 individuals over the age of 60 years and 126 patients with chronic liver disease were tested for HEV IgG. RESULTS: Forty cases of autochthonous hepatitis E (genotype 3) were identified. Hepatitis E was anicteric in 25% of cases and usually caused a self-limiting hepatitis predominantly in elderly Caucasian males. Six of 40 had a significant complication and three patients died, two of who had previously undiagnosed cirrhosis. Hepatitis E shows a seasonal variation with peaks in the spring and summer and no cases in November and December. HEV IgG prevalence increases with age, is more common in men and is 16% in blood donors, 13% in patients with chronic liver disease and 25% in individuals over 60 years. CONCLUSION: Autochthonous hepatitis E is more common than previously recognized, and should be considered in the differential diagnosis in patients with hepatitis, whatever their age or travel history. It carries a significant morbidity and when seen in the context of chronic liver disease carries an adverse prognosis.


Assuntos
Doadores de Sangue/estatística & dados numéricos , Hepatite E/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença Crônica , Inglaterra/epidemiologia , Feminino , Anticorpos Anti-Hepatite/sangue , Hepatite E/complicações , Hepatite E/diagnóstico , Hepatite E/imunologia , Vírus da Hepatite E/imunologia , Vírus da Hepatite E/isolamento & purificação , Humanos , Imunoglobulina G/sangue , Estilo de Vida , Masculino , Pessoa de Meia-Idade , Prognóstico , Estações do Ano , Estudos Soroepidemiológicos
6.
Vet Microbiol ; 126(1-3): 11-9, 2008 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-17656045

RESUMO

Six laboratories participated in a ring trial to evaluate the reliability of a real-time PCR assay for the detection of bovine herpesvirus 1 (BoHV-1) from extended bovine semen. Sets of coded samples were prepared and distributed to each of the laboratories. The sample panel contained semen from naturally and artificially infected bulls, serial dilutions of positive semen with negative semen, semen from uninfected seronegative bulls, negative semen spiked with virus, as well as serial dilutions of reference virus. The samples were tested using a previously validated real-time PCR assay for the detection of BoHV-1 in each participating laboratory. The PCR tests were conducted with four different real-time PCR amplification platforms, including RotorGene 3000, Stratagene MX 3000/4000, ABI 7900, and Roche LightCycler 2.0. Virus isolation using one set of samples was performed in one laboratory. The results of the laboratories were compared with one another, and with those of virus isolation. It was found that the sensitivity and specificity of the real-time PCR test was greater than those of virus isolation (82.7% versus 53.6% and 93.6% versus 84.6%, respectively). A high level of agreement on PCR testing results between the laboratories was achieved (kappa value 0.59-0.95). The results of this study indicate that the real-time PCR assay is suitable for the detection of BoHV-1 in extended semen, and would be a good substitute for the slow and laborious virus isolation, for the screening testing at artificial insemination centres and for international trade.


Assuntos
Herpesvirus Bovino 1/isolamento & purificação , Cooperação Internacional , Laboratórios , Reação em Cadeia da Polimerase/veterinária , Sêmen/virologia , Animais , Bovinos , Doenças dos Bovinos/virologia , Infecções por Herpesviridae/veterinária , Infecções por Herpesviridae/virologia , Herpesvirus Bovino 1/genética , Masculino , Variações Dependentes do Observador , Reprodutibilidade dos Testes , Preservação do Sêmen , Sensibilidade e Especificidade
7.
J Virol Methods ; 144(1-2): 103-8, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17561275

RESUMO

A real-time polymerase chain reaction (PCR) assay was developed for detection of the presence of bovine herpesvirus type 1 (BoHV-1) in extended bovine semen. The assay detects a region encoding a highly conserved glycoprotein B gene. The real-time PCR assay was validated for specificity, sensitivity and repeatability using spiked semen and semen from naturally infected animals. The real-time PCR was very rapid, highly repeatable and more sensitive (lower detection limits) than conventional virus isolation method for the detection of BoHV-1 in extended semen. The specificity of the assay is as expected. The assay had an analytical sensitivity of 0.38 TCID(50) virus spiked into negative semen. The second real-time PCR system for the detection of the bovine growth hormone (bGH) gene was applied as an internal control for the DNA extraction and PCR. The bGH PCR can be performed separately to BoHV-1 PCR, or in a duplex format. The real-time PCR assay is intended for use in international trade. The complete validation dossier based on this study and an international inter-laboratory ring trial has been accredited by the Office International des Epizooties (OIE) and has been recommended to be adopted as a prescribed test for international trade.


Assuntos
DNA Viral/análise , Herpesvirus Bovino 1/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Sêmen/virologia , Animais , Bovinos , Doenças dos Bovinos/virologia , Hormônio do Crescimento/análise , Hormônio do Crescimento/genética , Infecções por Herpesviridae/veterinária , Infecções por Herpesviridae/virologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
8.
Vet Immunol Immunopathol ; 119(3-4): 254-68, 2007 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-17629574

RESUMO

The aetiology of porcine post-weaning multisystemic wasting syndrome (PMWS) is poorly understood. Porcine circovirus type 2 (PCV-2) is an essential component of the experimental disease model for PMWS: however, evidence from experimental and field studies indicates that additional factors play a critical role in the aetiopathogenesis of PMWS. Current candidates include (1) immune stimulation (for example, via co-infection or vaccination), and (2) a novel infectious agent. A prospective, longitudinal case-control study was designed to investigate molecular triggers in leucocytes of neonatal piglets that may predispose to the development of PMWS. Blood samples were collected weekly from pigs (n=125) within five farms, from 1 week to 8 weeks of age: that is, before the appearance of clinical signs. Four colour flow cytometry was used to investigate changes in subsets of peripheral blood mononuclear cells, using monoclonal antibodies against the following cell associated markers; sIgG, CD3, MHCII dR, CD14, CD4a, CD8a, CD45RC, CD25, SWC3a, SWC8, CD163 and CD45. Sampling and laboratory analysis was supported by monitoring of clinical signs from 1 week to 20 weeks of age, or until disease supervened. At the conclusion of the study, 68 pigs (54%) were classified in Group 1 (no signs of clinical disease), 34 pigs (27%) in Group 2 (signs of clinical disease but not characteristic of PMWS), 17 pigs (14%) in Group 3 (suspect PMWS case) and 5 pigs (4%) in Group 4 (PMWS case). A single case of Porcine Dermatitis and Nephropathy syndrome (PDNS) was also diagnosed. Significant changes with age were demonstrated in clinically normal, neonatal pigs (Group 1), including an increase in B-cells and T-cells, and an increase in the proportion of total T-cells expressing MHCII. Within the T-cell subset, the proportion of CD8(+high) CD4(-) T-cells increased, in addition to the proportion of CD4(+) T-cells co-expressing CD8. Of the factors recorded, farm was found to have a highly significant effect on immune system development in the neonate. Comparison of Groups 1 and 4 cases identified significant differences between pigs which remained normal and those which subsequently developed PMWS. Pigs which went on to develop PMWS had a greater proportion of T-cells expressing MHCII in early life, higher mean intensity of expression of MHCII on T-cells, higher mean intensity of expression of MHCII on B cells and higher expression of CD25 on CD45RC(-) T-cells. These findings suggest that lymphocyte activation may be a key early event in the aetiology of PMWS.


Assuntos
Imunidade Celular/fisiologia , Síndrome Definhante Multissistêmico de Suínos Desmamados/imunologia , Suínos/crescimento & desenvolvimento , Suínos/imunologia , Envelhecimento , Animais , Animais Recém-Nascidos , Regulação da Expressão Gênica , Complexo Principal de Histocompatibilidade/genética , Complexo Principal de Histocompatibilidade/imunologia , Subpopulações de Linfócitos T/imunologia
9.
Vet Microbiol ; 96(4): 357-66, 2003 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-14599783

RESUMO

The aim of this study was to improve molecular methods for the detection of bovine viral diarrhoea virus (BVDV). A single-tube nested reverse-transcriptase polymerase chain reaction (nRT-PCR) employing the 5'-3'-exonuclease assay (TaqMan) system was optimised for use with bulk milk, semen and whole blood samples. An artificial template (mimic) was engineered to provide in-tube validation of negative samples by demonstrating the absence of substances inhibitory to RT or PCR. This mimic was constructed by disrupting the BVDV amplicon at the TaqMan probe site by inserting a 295bp fragment of human genomic DNA. The mimic amplicon was discriminated from the BVDV RT-PCR products using a second TaqMan probe, with a different fluorochrome specific for the inserted DNA. This new method was more sensitive than BVDV antigen ELISA methods and the existing RT-PCR method used in the laboratory for detection of BVDV in bulk milk. Furthermore, RNA extracted by robotic methods has proved suitable for use in this assay. This TaqMan nRT-PCR will be a valuable method for the detection of BVDV in a variety of biological matrices including milk and semen.


Assuntos
Doença das Mucosas por Vírus da Diarreia Viral Bovina/virologia , Vírus da Diarreia Viral Bovina/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Taq Polimerase/metabolismo , Animais , Anticorpos Antivirais/sangue , Bovinos , Vírus da Diarreia Viral Bovina/genética , Feminino , Corantes Fluorescentes/química , Masculino , Leite/virologia , Mutagênese Insercional , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sêmen/virologia , Sensibilidade e Especificidade , Taq Polimerase/genética , Moldes Genéticos
10.
Vet Microbiol ; 102(3-4): 169-81, 2004 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-15327792

RESUMO

Sets of serum and milk samples were collected from various countries and prepared, lyophilised and distributed by 1 laboratory to 12 reference laboratories in Europe. The serum sets contained the three European bovine herpesvirus 1 (BHV1) reference serum samples (EU1, EU2 and EU3), serum samples from naturally and experimentally BHV1-infected cattle, from vaccinated, and vaccinated-challenged cattle, from uninfected cattle, and a series of serum dilutions. In addition, sets of milk samples were distributed. The samples were tested for antibodies against BHV1 in virus neutralisation tests, in gB-specific ELISAs, in indirect ELISAs and in gE-specific ELISAs. It was found that the virus neutralisation test and the gB-specific ELISAs were most sensitive for the detection of antibodies in serum, whereas for assaying milk samples the indirect ELISAs were the tests of choice. The results show that the quality of most laboratories appeared to be adequate, but that one laboratory performed considerably below an acceptable level of quality. Four samples from the panel have been proposed that might be selected as reference sera in addition to the three European reference samples.


Assuntos
Anticorpos Antivirais/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/veterinária , Herpesvirus Bovino 1/imunologia , Rinotraqueíte Infecciosa Bovina/diagnóstico , Leite/imunologia , Testes de Neutralização/veterinária , Animais , Anticorpos Antivirais/análise , Anticorpos Antivirais/sangue , Bovinos , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Europa (Continente) , Laboratórios/normas , Leite/virologia , Testes de Neutralização/métodos , Testes de Neutralização/normas , Controle de Qualidade , Valores de Referência , Sensibilidade e Especificidade , Vacinação/veterinária , Proteínas Virais/imunologia , Vacinas Virais/imunologia
11.
J Wildl Dis ; 40(1): 99-102, 2004 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15137494

RESUMO

The aim of this study was to assess the frequency of mustelid herpesvirus-1 (MusHV-1) infection in free-ranging badgers (Meles meles) in the British Isles. A polymerase chain reaction assay was developed that detected MusHV-1 DNA in 95% (18/19) and 100% (10/10) of anticoagulant-treated blood samples collected from free-ranging badgers sampled in the southwest of England and the Republic of Ireland, respectively. An indirect immunoassay was also developed to detect MusHV-1-specific immunoglobulin-G in serum samples. Using an arbitrary cutoff of twice the optical density obtained with a virus-negative preparation, 32.7% (36/110) of sera sampled from badgers were positive. The conclusion drawn from these data is that infection with MusHV-1 is common among free-ranging badgers in the British Isles.


Assuntos
Carnívoros/virologia , Infecções por Herpesviridae/veterinária , Herpesviridae/isolamento & purificação , Reação em Cadeia da Polimerase/veterinária , Animais , Animais Selvagens/virologia , Anticorpos Antivirais/sangue , DNA Viral/análise , Herpesviridae/imunologia , Infecções por Herpesviridae/epidemiologia , Imunoensaio/veterinária , Irlanda/epidemiologia , Reação em Cadeia da Polimerase/métodos , Reino Unido/epidemiologia
13.
BMC Res Notes ; 5: 190, 2012 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-22534364

RESUMO

BACKGROUND: Hepatitis E virus (HEV) genotype 3 and 4 can cause liver disease in human and has its main reservoir in pigs. HEV investigations in pigs worldwide have been performed but there is still a lack of information on the infection dynamics in pig populations. FINDINGS: The HEV transmission dynamics in commercial pig farms in six different European countries was studied. The data collected show prevalence in weaners ranging from 8% to 30%. The average HEV prevalence in growers was between 20% and 44%. The fatteners prevalence ranged between 8% and 73%. Sows prevalence was similar in all countries. Boar faeces were tested for HEV only in Spain and Czech Republic, and the prevalence was 4.3% and 3.5% respectively. The collected data sets were analyzed using a recently developed model to estimate the transmission dynamics of HEV in the different countries confirming that HEV is endemic in pig farms. CONCLUSIONS: This study has been performed using similar detection methods (real time RT-PCR) for all samples and the same model (SIR model) to analyse the data. Furthermore, it describes HEV prevalence and within-herd transmission dynamics in European Countries (EU): Czech Republic, Italy, Portugal, Spain, The Netherlands and United Kingdom, confirming that HEV is circulating in pig farms from weaners to fatteners and that the reproductive number mathematical defined as R0 is in the same range for all countries studied.


Assuntos
Reservatórios de Doenças/veterinária , Vírus da Hepatite E/genética , Hepatite E/epidemiologia , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Doenças dos Suínos/epidemiologia , Fatores Etários , Animais , Europa (Continente)/epidemiologia , Fezes/virologia , Feminino , Genótipo , Hepatite E/transmissão , Hepatite E/virologia , Vírus da Hepatite E/classificação , Vírus da Hepatite E/isolamento & purificação , Humanos , Masculino , Filogenia , Prevalência , RNA Viral/classificação , RNA Viral/isolamento & purificação , Suínos , Doenças dos Suínos/transmissão
16.
J Infect Dis ; 192(7): 1166-72, 2005 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-16136458

RESUMO

Between 1996 and 2003, 186 cases of hepatitis E were serologically diagnosed. Of these, 17 (9%) were not associated with recent travel abroad. Patients were >55 years old (range, 56-82 years old) and tended to be male (76%). Two patients presented with fulminant hepatitis. A total of 129 (69%) cases were associated with recent travel to countries where hepatitis E virus (HEV) is hyperendemic. Compared with patients with travel-associated disease, patients with non-travel-associated disease were more likely to be older, living in coastal or estuarine areas, not of South Asian ethnicity, and infected by genotype 3 strains of HEV. The genotype 3 subgenomic nucleotide sequences were unique and closely related to those from British pigs. Patients infected by HEV indigenous to England and Wales tended to belong to a distinct demographic group, there were multiple sources of infection, and pigs might have been a viral reservoir.


Assuntos
Vírus da Hepatite E/genética , Hepatite E/epidemiologia , Hepatite E/fisiopatologia , Epidemiologia Molecular , Viagem , Idoso , Idoso de 80 Anos ou mais , Demografia , Inglaterra/epidemiologia , Feminino , Anticorpos Anti-Hepatite/sangue , Hepatite E/virologia , Vírus da Hepatite E/classificação , Vírus da Hepatite E/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , País de Gales/epidemiologia
17.
Emerg Infect Dis ; 10(5): 953-5, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15200841

RESUMO

We describe a case of acquired infection of a strain of hepatitis E virus (HEV) with a 100% amino acid identity to the analogous region in strains of HEV circulating in a United Kingdom pig herd. This case further supports the theory that autochthonous HEV infection in industrialized countries is zoonotic.


Assuntos
Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Hepatite E/virologia , Doenças dos Suínos/virologia , Zoonoses/virologia , Animais , Feminino , Hepatite E/veterinária , Vírus da Hepatite E/isolamento & purificação , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Análise de Sequência de DNA , Suínos , Reino Unido
18.
J Gen Virol ; 83(Pt 6): 1325-1330, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12029147

RESUMO

A herpesvirus causing a cytopathic effect was isolated from pulmonary fibroblast cultures established from a European badger (Meles meles). A study was undertaken to classify and to assess some in-vitro growth characteristics of this virus. From a panel of 27 mammalian cell lines, in-vitro replication of the badger herpesvirus (BadHV) was only demonstrated with a mink lung cell line, suggesting a high degree of host specificity. Using PCR with degenerate primers, three independent fragments of the BadHV genome were sequenced. The largest of these fragments comprised a 6.2 kb segment including the DNA polymerase and glycoprotein B genes. Phylogenetic analysis of these sequences demonstrated that the BadHV is novel and clearly grouped with members of the Gammaherpesvirinae. In view of the oncogenic and immunosuppressive potential of many related herpesviruses, it is possible that BadHV can impact on existing acute or chronic disease in badgers.


Assuntos
Carnívoros/virologia , Gammaherpesvirinae/isolamento & purificação , Genoma Viral , Sequência de Aminoácidos , Animais , Linhagem Celular , DNA Polimerase Dirigida por DNA/genética , Europa (Continente) , Gammaherpesvirinae/classificação , Gammaherpesvirinae/genética , Glicoproteínas/genética , Pulmão , Vison , Dados de Sequência Molecular , Alinhamento de Sequência , Especificidade da Espécie , Proteínas Virais/genética
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