RESUMO
African horse sickness virus serotype 9 (AHSV-9) has been known for some time to be circulating amongst equids in West Africa without causing any clinical disease in indigenous horse populations. Whether this is due to local breeds of horses being resistant to disease or whether the AHSV-9 strains circulating are avirulent is currently unknown. This study shows that the majority (96%) of horses and donkeys sampled across The Gambia were seropositive for AHS, despite most being unvaccinated and having no previous history of showing clinical signs of AHS. Most young horses (<3 years) were seropositive with neutralizing antibodies specific to AHSV-9. Eight young equids (<3 years) were positive for AHSV-9 by serotype-specific RT-PCR and live AHSV-9 was isolated from two of these horses. Sequence analysis revealed the presence of an AHSV-9 strain showing 100% identity to Seg-2 of the AHSV-9 reference strain, indicating that the virus circulating in The Gambia was highly likely to have been derived from a live-attenuated AHSV-9 vaccine strain.
Assuntos
Vírus da Doença Equina Africana/isolamento & purificação , Doença Equina Africana/epidemiologia , Anticorpos Antivirais/sangue , Vacinas Virais , Vírus da Doença Equina Africana/classificação , Vírus da Doença Equina Africana/imunologia , Animais , Anticorpos Neutralizantes/sangue , Equidae , Gâmbia/epidemiologia , Cavalos , RNA Viral/genética , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência , Estudos Soroepidemiológicos , Sorotipagem , Vacinas AtenuadasRESUMO
Prior to the recent outbreak of equine encephalosis in Israel in 2009, equine encephalosis virus (EEV) had only been isolated from equids in South Africa. In this study we show the first evidence for the circulation of EEV beyond South Africa in Ethiopia, Ghana and The Gambia, indicating that EEV is likely to be freely circulating and endemic in East and West Africa. Sequence analysis revealed that the EEV isolate circulating in The Gambia was closely related to an EEV isolate that was isolated from a horse from Israel during the EEV outbreak in 2009, indicating that the two viruses have a common ancestry. Interestingly horses in Morocco tested negative for EEV antibodies indicating that the Sahara desert may be acting as a geographical barrier to the spread to the virus to North African countries. This evidence for EEV circulation in countries in East and West Africa sheds light on how the virus may have reached Israel to cause the recent outbreak in 2009.
Assuntos
Doenças dos Cavalos/epidemiologia , Orbivirus/isolamento & purificação , Infecções por Reoviridae/veterinária , Animais , Anticorpos Antivirais/sangue , Sequência de Bases , Surtos de Doenças/veterinária , Ensaio de Imunoadsorção Enzimática , Equidae , Etiópia/epidemiologia , Gâmbia/epidemiologia , Gana/epidemiologia , Doenças dos Cavalos/virologia , Cavalos , Israel/epidemiologia , Dados de Sequência Molecular , Orbivirus/classificação , Orbivirus/genética , Orbivirus/imunologia , Filogenia , RNA Viral , Infecções por Reoviridae/epidemiologia , Infecções por Reoviridae/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estudos Soroepidemiológicos , SorotipagemRESUMO
CD8+ cytotoxic T lymphocytes (CTL) can be effective at controlling HIV-1 in humans and SIV in macaques, but their utility is partly offset by mutational escape. The kinetics of CTL escape and reversion of escape mutant viruses upon transmission to MHC-mismatched hosts can help us understand CTL-mediated viral control and the fitness cost extracted by immune escape mutation. Traditional methods for following CTL escape and reversion are, however, insensitive to minor viral quasispecies. We developed sensitive quantitative real-time PCR assays to track the viral load of SIV Gag164-172 KP9 wild-type (WT) and escape mutant (EM) variants in pigtail macaques. Rapid outgrowth of EM virus occurs during the first few weeks of infection. However, the rate of escape plateaued soon after, revealing a prolonged persistence of WT viremia not detectable by standard cloning and sequencing methods. The rate of escape of KP9 correlated with levels of vaccine-primed KP9-specific CD8+ T cells present at that time. Similarly, when non-KP9 responder (lacking the restricting Mane-A*10 allele) macaques were infected with SHIVmn229 stock containing a mixture of EM and WT virus, rapid reversion to WT was observed over the first 2 weeks following infection. However, the rate of reversion to WT slowed dramatically over the first month of infection. The serial quantitation of escape mutant viruses evolving during SIV infection shows that rapid dynamics of immune escape and reversion can be observed in early infection, particularly when CD8 T cells are primed by vaccination. However, these early rapid rates of escape and reversion are transient and followed by a significant slowing in these rates later during infection, highlighting that the rate of escape is significantly influenced by the timing of its occurrence.
Assuntos
Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/imunologia , Vacinação , Doença Aguda , Animais , Sequência de Bases , Doença Crônica , Epitopos de Linfócito T/imunologia , Imunidade Celular/imunologia , Macaca nemestrina , Dados de Sequência Molecular , Mutação , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Síndrome de Imunodeficiência Adquirida dos Símios/sangue , Vírus da Imunodeficiência Símia/genética , Linfócitos T Citotóxicos/imunologia , Fatores de TempoRESUMO
Effective immunotherapies for HIV are needed. Drug therapies are life-long with significant toxicities. Dendritic-cell based immunotherapy approaches are promising but impractical for widespread use. A simple immunotherapy, reinfusing fresh autologous blood cells exposed to overlapping SIV peptides for 1 hour ex vivo, was assessed for the control of SIV(mac251) replication in 36 pigtail macaques. An initial set of four immunizations was administered under antiretroviral cover and a booster set of three immunizations administered 6 months later. Vaccinated animals were randomized to receive Gag peptides alone or peptides spanning all nine SIV proteins. High-level, SIV-specific CD4 and CD8 T-cell immunity was induced following immunization, both during antiretroviral cover and without. Virus levels were durably approximately 10-fold lower for 1 year in immunized animals compared to controls, and a significant delay in AIDS-related mortality resulted. Broader immunity resulted following immunizations with peptides spanning all nine SIV proteins, but the responses to Gag were weaker in comparison to animals only immunized with Gag. No difference in viral outcome occurred in animals immunized with all SIV proteins compared to animals immunized against Gag alone. Peptide-pulsed blood cells are an immunogenic and effective immunotherapy in SIV-infected macaques. Our results suggest Gag alone is an effective antigen for T-cell immunotherapy. Fresh blood cells pulsed with overlapping Gag peptides is proceeding into trials in HIV-infected humans.
Assuntos
Produtos do Gene gag/administração & dosagem , Imunoterapia/métodos , Síndrome de Imunodeficiência Adquirida dos Símios/prevenção & controle , Vírus da Imunodeficiência Símia/imunologia , Viremia/terapia , Animais , Transfusão de Componentes Sanguíneos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Modelos Animais de Doenças , Feminino , Produtos do Gene gag/imunologia , Longevidade , Macaca nemestrina , Masculino , Síndrome de Imunodeficiência Adquirida dos Símios/imunologiaRESUMO
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has dramatically expedited global vaccine development efforts1-3, most targeting the viral 'spike' glycoprotein (S). S localizes on the virion surface and mediates recognition of cellular receptor angiotensin-converting enzyme 2 (ACE2)4-6. Eliciting neutralizing antibodies that block S-ACE2 interaction7-9, or indirectly prevent membrane fusion10, constitute an attractive modality for vaccine-elicited protection11. However, although prototypic S-based vaccines show promise in animal models12-14, the immunogenic properties of S in humans are poorly resolved. In this study, we characterized humoral and circulating follicular helper T cell (cTFH) immunity against spike in recovered patients with coronavirus disease 2019 (COVID-19). We found that S-specific antibodies, memory B cells and cTFH are consistently elicited after SARS-CoV-2 infection, demarking robust humoral immunity and positively associated with plasma neutralizing activity. Comparatively low frequencies of B cells or cTFH specific for the receptor binding domain of S were elicited. Notably, the phenotype of S-specific cTFH differentiated subjects with potent neutralizing responses, providing a potential biomarker of potency for S-based vaccines entering the clinic. Overall, although patients who recovered from COVID-19 displayed multiple hallmarks of effective immune recognition of S, the wide spectrum of neutralizing activity observed suggests that vaccines might require strategies to selectively target the most potent neutralizing epitopes.
Assuntos
Anticorpos Neutralizantes/farmacologia , Infecções por Coronavirus/imunologia , Peptidil Dipeptidase A/genética , Pneumonia Viral/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Enzima de Conversão de Angiotensina 2 , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/farmacologia , Antígenos Virais/imunologia , COVID-19 , Chlorocebus aethiops , Infecções por Coronavirus/patologia , Infecções por Coronavirus/virologia , Epitopos/imunologia , Humanos , Imunidade Celular/imunologia , Pandemias , Peptidil Dipeptidase A/imunologia , Pneumonia Viral/patologia , Pneumonia Viral/virologia , Glicoproteína da Espícula de Coronavírus/antagonistas & inibidores , Linfócitos T Auxiliares-Indutores/imunologia , Células Vero/imunologiaRESUMO
Bluetongue virus European Community national reference laboratories (BTV-EC-NRLs) participated in an inter-laboratory proficiency test in 2007. The aim of the inter-laboratory proficiency test was to determine the ability of laboratories to detect antibodies to a series of BTV serotypes by cELISA and to detect viral RNA in animals infected with the European strain of BTV-8 by RT-PCR. Both serum and EDTA blood sample were diluted in order to determine the sensitivity of the assays. All the cELISAs were 'fit-for purpose' to detect antibodies to the common BTV serotypes circulating in Europe and the real time RT-PCR assays were all capable of detecting BTV-8 RNA albeit with varying sensitivities. There were however inconsistencies in the ability of the gel-based PCR assays to detect BTV RNA. In addition, samples taken on the first day of viraemia and at the peak of viraemia from animals experimentally infected with BTV-8, were diluted to determine if the diluting of samples affected the ability of the Shaw et al. (Shaw, A.E., M., P., Alpar, H.O., Anthony, S., Darpel, K.E., Batten, C.A., Carpenter, S., Jones, H., Oura, C.A.L., King, D.P., Elliott, H., Mellor, P.S., Mertens, P.P.C., 2007. Development and validation of a real-time RT-PCR assay to detect genome bluetongue virus segment 1. Journal of Virological Methods) RT-PCR assay to detect BTV-RNA at these time-points. Results indicated that, if samples were taken at the onset of viraemia, diluting at 1/5 resulted in a reduced ability of the assay to detect BTV RNA in the diluted compared to the neat samples. Diluting samples taken at the peak of viraemia at 1/10 however resulted in no loss in sensitivity.
Assuntos
Vírus Bluetongue/isolamento & purificação , Bluetongue/epidemiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Animais , Bluetongue/sangue , Bluetongue/diagnóstico , Vírus Bluetongue/genética , Ensaio de Imunoadsorção Enzimática , Europa (Continente) , União Europeia , RNA Viral/genética , RNA Viral/isolamento & purificação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Ovinos , Viremia/diagnóstico , Viremia/veterináriaRESUMO
European Community national reference laboratories participated in two inter-laboratory comparison tests in 2006 to evaluate the sensitivity and specificity of their 'in-house' ELISA and RT-PCR assays for the detection of bluetongue virus (BTV) antibodies and RNA. The first ring trial determined the ability of laboratories to detect antibodies to all 24 serotypes of BTV. The second ring trial, which included both antisera and EDTA blood samples from animals experimentally infected with the northern European strain of BTV-8, determined the ability of laboratories to detect BTV-8 antibodies and RNA, as well as the diagnostic sensitivity of the assays. A total of six C-ELISAs, six real-time RT-PCR and three conventional RT-PCR assays were used. All C-ELISAs were capable of detecting the BTV serotypes currently circulating in Europe (BTV-1, 2, 4, 8, 9 and 16), however some assays displayed inconsistencies in the detection of other serotypes, particularly BTV-19. All C-ELISAs detected BTV-8 antibodies in cattle and sheep by 21 dpi, while the majority of assays detected antibodies by 9 dpi in cattle and 8 dpi in sheep. All the RT-PCR assays were able to detect BTV-8, although the real-time assays were more sensitive compared to the conventional assays. The majority of real-time RT-PCR assays detected BTV RNA as early as 2 dpi in cattle and 3 dpi in sheep. These two ring trails provide evidence that national reference laboratories within the EC are capable of detecting BTV antibodies and RNA and provide specificity and sensitivity information on the detection methods currently available.
Assuntos
Vírus Bluetongue/isolamento & purificação , Bluetongue/diagnóstico , Ensaio de Imunoadsorção Enzimática/veterinária , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Animais , Anticorpos Antivirais/sangue , Vírus Bluetongue/genética , Vírus Bluetongue/imunologia , Bovinos , DNA Viral/sangue , União Europeia , Distribuição Aleatória , OvinosRESUMO
This paper presents evidence that a field strain of bluetongue virus serotype 8 (BTV-8) was transmitted transplacentally and that it was also spread by a direct contact route. Twenty pregnant heifers were imported from the Netherlands into Northern Ireland during the midge-free season. Tests before and after the animals were imported showed that eight of them had antibodies to bluetongue virus, but no viral RNA was detected in any of them by reverse transcriptase-PCR (RT-PCR). Two of the seropositive heifers gave birth to three calves that showed evidence of bluetongue virus infection (RT-PCR-positive), and one of the calves was viraemic. Two further viraemic animals (one newly calved Dutch heifer, and one milking cow originally from Scotland) were also found to have been infected with BTV-8 and evidence is presented that these two animals may have been infected by direct contact, possibly through the ingestion of placentas infected with BTV-8.
Assuntos
Bluetongue/transmissão , Doenças dos Bovinos/transmissão , Transmissão Vertical de Doenças Infecciosas/veterinária , Animais , Animais Recém-Nascidos/virologia , Anticorpos Antivirais/isolamento & purificação , Bluetongue/epidemiologia , Vírus Bluetongue/isolamento & purificação , Bovinos , Doenças dos Bovinos/virologia , Ceratopogonidae , Surtos de Doenças/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Insetos Vetores , Entrevistas como Assunto , Masculino , Países Baixos , Irlanda do Norte/epidemiologia , Placenta/virologia , Gravidez , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sorotipagem/veterinária , OvinosRESUMO
In 2014, a new bluetongue virus serotype 4 (BTV-4) strain was detected in southern Greece and spread rapidly throughout the Balkan Peninsula and adjacent countries. Within half a year, more than 7,068 outbreaks were reported in ruminants, particularly in sheep. However, the reported morbidity and case fatality rates in ruminants varied. The pathogenesis of a Bulgarian BTV-4 strain isolated from sheep during the BTV-4 epizootic was studied in different species. Therefore, four sheep, three goats and three cattle were experimentally infected with the isolate BTV-4/BUL2014/15 and monitored for clinical signs up to several weeks. Serum and whole-blood samples were collected at regular intervals and subjected to serological and virological analyses. In this context, BTV-4-specific real-time RT-PCR assays were developed. The infection kinetics were similar to those known for other traditional BTV serotypes, and only mild BT-like clinical signs were observed in goats and sheep. In cattle, no obvious clinical signs were observed, except a transient increase in body temperature. The study results contrast with the severe clinical signs reported in sheep experimentally infected with an African BTV-4 strain and with the reports of BT-like clinical signs in a considerable proportion of different ruminant species infected with BTV-4 in the Balkan region and Italy. The discrepancies between the results of these animal trials and observations of BTV-4 infection in the field may be explained by the influence of various factors on the manifestation of BT disease, such as animal breed, fitness and virus strain, as described previously.
Assuntos
Vírus Bluetongue/patogenicidade , Bluetongue/virologia , Doenças dos Bovinos/virologia , Surtos de Doenças/veterinária , Doenças das Cabras/virologia , Animais , Bluetongue/epidemiologia , Vírus Bluetongue/imunologia , Vírus Bluetongue/isolamento & purificação , Bulgária/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças das Cabras/epidemiologia , Cabras , Masculino , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Sorogrupo , OvinosRESUMO
The European Virus Archive (EVA) was created in 2008 with funding from the FP7-EU Infrastructure Programme, in response to the need for a coordinated and readily accessible collection of viruses that could be made available to academia, public health organisations and industry. Within three years, it developed from a consortium of nine European laboratories to encompass associated partners in Africa, Russia, China, Turkey, Germany and Italy. In 2014, the H2020 Research and Innovation Framework Programme (INFRAS projects) provided support for the transformation of the EVA from a European to a global organization (EVAg). The EVAg now operates as a non-profit consortium, with 26 partners and 20 associated partners from 21 EU and non-EU countries. In this paper, we outline the structure, management and goals of the EVAg, to bring to the attention of researchers the wealth of products it can provide and to illustrate how end-users can gain access to these resources. Organisations or individuals who would like to be considered as contributors are invited to contact the EVAg coordinator, Jean-Louis Romette, at jean-louis.romette@univmed.fr.
Assuntos
Arquivos , Bancos de Espécimes Biológicos/organização & administração , Recursos em Saúde/organização & administração , Vírus , Pesquisa Biomédica , Europa (Continente) , Humanos , Disseminação de Informação , Organizações de Serviços Gerenciais , Coronavírus da Síndrome Respiratória do Oriente Médio , Saúde Pública , Controle de Qualidade , Segurança/normas , Virologia/métodos , Febre Amarela/epidemiologia , Febre Amarela/virologia , Infecção por Zika virus/epidemiologia , Infecção por Zika virus/virologiaRESUMO
Since 1998, multiple strains of bluetongue virus (BTV), belonging to six different serotypes (types 1, 2, 4, 8, 9 and 16) have caused outbreaks of disease in Europe, causing one of the largest epizootics of bluetongue ever recorded, with the deaths of >1.8 million animals (mainly sheep). The persistence and continuing spread of BTV in Europe and elsewhere highlights the importance of sensitive and reliable diagnostic assay systems that can be used to rapidly identify infected animals, helping to combat spread of the virus and disease. BTV has a genome composed of 10 linear segments of dsRNA. We describe a real-time RT-PCR assay that targets the highly conserved genome segment 1 (encoding the viral polymerase--VP1) that can be used to detect all of the 24 serotypes, as well as geographic variants (different topotypes) within individual serotypes of BTV. After an initial evaluation using 132 BTV samples including representatives of all 24 BTV serotypes, this assay was used by the European Community Reference Laboratory (CRL) at IAH Pirbright to confirm the negative status of 2,255 animals imported to the UK from regions that were considered to be at risk during the 2006 outbreak of BTV-8 in Northern Europe. All of these animals were also negative by competition ELISA to detect BTV specific antibodies and none of them developed clinical signs of infection. These studies have demonstrated the value of the assay for the rapid screening of field samples.
Assuntos
Vírus Bluetongue/isolamento & purificação , Genoma Viral , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sequência de Aminoácidos , Animais , Bluetongue/diagnóstico , Vírus Bluetongue/classificação , Vírus Bluetongue/genética , Dados de Sequência Molecular , RNA Viral/isolamento & purificação , Sensibilidade e Especificidade , Alinhamento de Sequência , Carneiro DomésticoRESUMO
Four poll Dorset sheep and four Holstein-Friesian cattle were infected with the northern European strain of bluetongue virus (BTV), BTV-8, to assess its pathogenicity in UK breeds. The time course of infection was monitored in both species by using real-time reverse transcriptase-PCR (RT-PCR), conventional RT-PCR and serology. Two of the sheep developed severe clinical signs that would have been fatal in the field; the other two were moderately and mildly ill, respectively. The cattle were clinically unaffected, but had high levels of viral RNA in their bloodstream. Real-time RT-PCR detected viral RNA as early as one day after infection in the cattle and three days after infection in the sheep. Antibodies against BTV were detected by six days after infection in the sheep and eight days after infection in the cattle. Postmortem examinations revealed pathology in the cattle that was more severe than suggested by the mild clinical signs, but the pathological and clinical findings in the sheep were more consistent.
Assuntos
Vírus Bluetongue/patogenicidade , Bluetongue , Doenças dos Bovinos/patologia , Doenças dos Ovinos/patologia , Animais , Bluetongue/epidemiologia , Bluetongue/patologia , Bluetongue/virologia , Vírus Bluetongue/isolamento & purificação , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Surtos de Doenças/veterinária , Europa (Continente)/epidemiologia , Feminino , Masculino , RNA Viral/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Índice de Gravidade de Doença , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/virologia , Especificidade da Espécie , Reino Unido/epidemiologia , Carga Viral/veterináriaRESUMO
African horse sickness (AHS) is a disease of equids caused by African Horse Sickness Virus (AHSV) and is transmitted by Culicoides midges. AHS is endemic in sub-Saharan Africa, but during the past century, outbreaks of significant economic importance and elevated mortality have been recorded in Northern African countries, the Iberian and Arabian Peninsula, the Middle East and the Indian subcontinent. Effective control combines the application of early warning systems, accurate laboratory diagnosis and reporting, animal movement restrictions, suitable vaccination and surveillance programs, and the coordination of all these measures by efficient veterinary services. Conventional reverse-transcriptase (RT) PCR (RT-PCR) and real-time RT-PCR (rRT-PCR) assays have improved the sensitivity and rapidity of diagnosing AHS, resulting in the adoption of these methods as recommended tests by the World Organisation for Animal Health (OIE). However, currently these assays are only performed within laboratory settings; therefore, the development of field diagnostics for AHS would improve the fast implementation of control policies. Loop-mediated isothermal amplification (LAMP) is an isothermal, autocycling, strand-displacement nucleic acid amplification technique which can be performed in the field. LAMP assays are attractive molecular assays because they are simple to use, rapid, portable and have sensitivity and specificity within the range of rRT-PCR. This study describes the development of a novel RT-LAMP assay for the detection of AHSV. The AHSV RT-LAMP assay has an analytical sensitivity of 96.1% when considering an rRT-PCR cut-off value of CT > 36, or 91.3% when no rRT-PCR cut-off is applied. Diagnostic sensitivity and specificity were 100%. This assay provides for a rapid and low cost AHS diagnostic for use in the field.
Assuntos
Vírus da Doença Equina Africana/isolamento & purificação , Doença Equina Africana/diagnóstico , Ceratopogonidae/virologia , Técnicas de Amplificação de Ácido Nucleico/veterinária , Doença Equina Africana/virologia , Vírus da Doença Equina Africana/genética , Animais , Cavalos , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e EspecificidadeRESUMO
To better understand risks associated with trading cattle, it is important to know which serotypes of Bluetongue virus (BTV) are circulating within the exporting and importing country. Hence, this study was conducted to identify the circulating serotypes of BTV in Trinidad. Blood samples were collected monthly from sixty BTV- naïve imported cattle over a six month period after their arrival in the country. Virological (PCR and virus isolation) and serological (ELISA) analyses were carried out on the samples and CDC light traps were placed near the cattle enclosure to trap and identify the species of Culicoides biting midges that were present. All of the cattle seroconverted for BTV antibodies within three months of their arrival in the country and real-time reverse transcription PCR (rRT-PCR) detected BTV-RNA in the samples throughout the remainder of the study. The patterns of infection observed in the cattle indicated serial infections with multiple serotypes. A combination of BTV serotype-specific rRT-PCR on the original blood samples and virus isolation followed by serotype-specific rRT-PCR on selected samples, confirmed the presence of BTV serotypes 1, 2, 3, 5, 12 and 17. This is the first report of BTV-2 and BTV-5 in Trinidad. Light-suction traps placed in close proximity to the cattle predominantly trapped Culicoides insignis Lutz 1913 species (96%), with a further six Culicoides species making up the remaining 4% of trapped samples. The circulation of multiple BTV serotypes in Trinidad underlines the need for regular surveillance, which will contribute to the development of risk assessments for trade in livestock.
Assuntos
Anticorpos Antivirais/sangue , Vírus Bluetongue/imunologia , Bluetongue/virologia , Doenças dos Bovinos/virologia , Ceratopogonidae/virologia , Insetos Vetores/virologia , Animais , Bluetongue/epidemiologia , Vírus Bluetongue/isolamento & purificação , Bovinos , Doenças dos Bovinos/epidemiologia , Feminino , Masculino , Sorogrupo , Trinidad e Tobago/epidemiologiaRESUMO
The global impact of HIV/AIDS intensifies the need for a preventive vaccine and nonhuman primate models can help provide critical insights into effective immunity. Pigtail macaques (Macaca nemestrina) are increasingly studied as a nonhuman primate model for AIDS. We compared the virologic and immunologic characteristics of HIV-1, SIV, and SHIV infection of naive pigtail macaques across a series of preclinical HIV vaccine studies. SIVmac251 and SIVmac239 infection of naive pigtail macaques resulted in a gradual decline in peripheral CD4+ T cells in the setting of high levels of viremia, approximating most closely human infection of HIV-1. In contrast, the CXCR4-utilizing SHIVmn229 virus resulted in rapid depletion of CD4+ T cells and minimal generation of humoral or cellular immune responses, similar to that observed with SHIV89.6P infection of rhesus macaques. Infection with the CCR5-utilizing, rhesus macaque passaged, SHIVSF162P3 resulted in some overall CD4+ T cell decline, however, three of eight macaques naturally control SHIVSF162P3 viremia to very low levels in the setting of robust adaptive immunity. Despite attempts at infecting pigtail macaques with HIV-1 strains passaged in juvenile pigtail macaques in vivo or in PBMC isolated from pigtail macaques in vitro, only lower nonsustained levels of viral replication were observed. Our results provide a series of virologic models with which to evaluate potential AIDS vaccines in pigtail macaques.
Assuntos
Modelos Animais de Doenças , Infecções por HIV/fisiopatologia , HIV-1/patogenicidade , Vírus da Imunodeficiência Símia/patogenicidade , Animais , Anticorpos Antivirais/sangue , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Anticorpos Anti-HIV/sangue , Infecções por HIV/imunologia , Infecções por HIV/virologia , Humanos , Macaca nemestrina , Testes de Neutralização , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/fisiopatologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologiaRESUMO
Isolates of peste des petits ruminants virus (PPRV) can be segregated genetically into four lineages. For decades, lineages I-III have been reported across Africa whilst lineage IV has predominantly circulated across Asia. However, the lineage distribution is currently changing in Africa. Importantly, full genome sequence data for African field isolates have been lacking. Here, we announce the first complete genome sequence of a field isolate of peste des petits ruminants virus (PPRV) from East Africa. This isolate was derived from the intestine of a goat suffering from severe clinical disease during the 2010 outbreak in Ethiopia. The full genome sequence of this isolate, PPRV Ethiopia/2010, clusters genetically with other lineage IV isolates of PPRV, sharing high levels of sequence identity across the genome. Further, we have carried out a phylogenetic analysis of all of the available African partial N gene and F gene PPRV sequences to investigate the epidemiology of PPRV with a focus on the emergence of different lineages of PPRV in Africa.
Assuntos
Peste dos Pequenos Ruminantes/epidemiologia , Vírus da Peste dos Pequenos Ruminantes/genética , Animais , Etiópia/epidemiologia , Genoma Viral , Filogenia , Análise de Sequência de DNARESUMO
The effects of various hormones on the urinary excretion of kallikrein and esterase A2 were studied in rats. Chronic treatment with antidiuretic hormone had no effect on the excretion of either enzyme. Deoxycorticosterone treatment or a low sodium diet stimulated urinary kallikrein excretion (as is well known), but had no effect on urinary esterase A2. Dexamethasone markedly suppressed the excretion of both kallikrein and esterase A2 and increased the excretion of proteins (inhibitors) that bind to each of these enzymes. It is likely that the suppressive effect of glucocorticoid on urinary kallikrein and esterase A2 activities is due to the increase in urinary binding proteins. There is also a strong correlation between the excretion of kallikrein and esterase A2 in normal untreated rats. Such a correlation might arise from a common effect of glucocorticoid-influenced inhibitors on both enzymes.
Assuntos
Aprotinina/urina , Calicreínas/urina , Peptídeo Hidrolases/urina , Animais , Dexametasona/farmacologia , Dieta Hipossódica , Feminino , Inibidores de Proteases/urina , Ratos , Ratos Endogâmicos , Vasopressinas/farmacologiaRESUMO
Dahl salt-sensitive (S) rats which are susceptible to hypertension have lower urinary kallikrein excretion than salt-resistant (R) rats which are not susceptible. Some physicochemical characteristics of partially purified urinary kallikrein were compared between the S and R strains. The isoelectric focusing pattern of S kallikrein was shifted so that a higher proportion of enzyme was present in isoelectric forms that had higher pI values compared to the pattern for R kallikrein. This strain difference was unique to urinary kallikrein; it was not seen in kallikrein extracted from salivary glands. The isoelectric focusing pattern for R urinary kallikrein could be converted to an S-type pattern by treatment with neuraminidase, which suggests that the differing isoelectric focusing patterns arose from differences in the sialic acid content of the kallikrein. The S kallikrein was slightly more heat-labile than R kallikrein, which was also compatible with the lower sialic acid content of the S enzyme. Tests involving the active site of the enzyme (Km values, pH curves, and heat of activation) were identical for the S and R strains. It was concluded that the structural differences observed in urinary kallikrein between S and R strains were compatible with strain-specific posttranslational processing of the enzyme.
Assuntos
Hipertensão/genética , Calicreínas/urina , Ratos Endogâmicos/genética , Cloreto de Sódio/fisiologia , Animais , Suscetibilidade a Doenças , Hipertensão/induzido quimicamente , Hipertensão/urina , Focalização Isoelétrica , Calicreínas/metabolismo , Cinética , Neuraminidase/urina , Peptídeo Hidrolases/metabolismo , Ratos , Ratos Endogâmicos/metabolismo , Glândulas Salivares/metabolismoRESUMO
The occurrence of hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase in adult Hymenolepis diminuta was demonstrated. This activity was negligible in the cestode's cytosolic fraction but was noted when the mitochondrial or microsomal fraction served as the enzyme source. The predominant localization of HMG-CoA reductase activity was with the microsomal fraction. This fraction did not contain appreciable mitochondrial contamination based on the distribution of marker enzymes. The enzymatic nature of HMG-CoA conversion to mevalonic acid by either fraction was apparent because the reaction was heat labile and responded linearly to time of assay and protein content. The enzymatic reduction of HMG-CoA absolutely required NADPH when either fraction was assayed. The lesser activity of the mitochondrial fraction was membrane-associated. The predominant localization of HMG-CoA reductase activity with microsomal membranes and its separation with the membranous component of the mitochondrial fraction suggest that mitochondrial activity reflects the presence of microsomal membranes. In its predominant localization and pyridine nucleotide requirement, the cestode's HMG-CoA reductase activity resembles that of mammalian systems. The finding of HMG-CoA reductase provides an enzymatic mechanism for the intermediate conversion of HMG-CoA to mevalonic acid that would be needed for acetate-dependent isoprenoid lipid synthesis by adult H. diminuta.
Assuntos
Hidroximetilglutaril-CoA Redutases/metabolismo , Hymenolepis/enzimologia , Animais , Ácido Mevalônico/análise , Ácido Mevalônico/metabolismo , Microssomos/enzimologia , Mitocôndrias/enzimologia , NADP/metabolismo , Succinato Desidrogenase/análiseRESUMO
OBJECTIVES: To study the epidemiological characteristics of patients with chronic hepatitis C virus followed in a primary referral hospital and the clinical influence of "systematic screening" defined as the screening of patients without symptoms and with known risk factors of hepatitis C (past transfusion, past or present intravenous drug use, haemodialysis) on the natural history and treatment of chronic hepatitis C virus. METHODS: The files of 311 consecutive patients who screened positive for anti-hepatitis C virus and were seen at the primary referral hospital, Creil, from January 1992 to February 1996, were analyzed. RESULTS: Patients who underwent "systematic screening" were younger with a shorter duration of infection. They were more often intravenous drug addicts and had lower alanine aminotransferase activity and Knodell scores than patients who underwent screening during "a diagnostic procedure", because of symptoms and/or abnormal liver biochemistry. Increased age at contamination and alcohol consumption of more than 40 g per day was associated with an increased risk of cirrhosis while patients who underwent "systematic screening" had a lower risk of cirrhosis and higher survival rate. Interferon therapy was attempted less often in anti-hepatitis C virus positive patients from "systematic screening" programs. CONCLUSIONS: Anti-hepatitis C virus positive patients from "systematic screening" programs had a benign disease and were rarely treated with interferon compared to anti-hepatitis C virus positive patients diagnosed during a "diagnostic procedure".