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1.
Trop Anim Health Prod ; 50(1): 43-48, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29022241

RESUMO

The caprine arthrite encephalite (CAE) is a disease that affects especially dairy goat. The virus shows compartmentalization features, that allows it to hide at certain times during the course of the disease, making it difficult to control. The present study was conducted to identify the major seminal plasma protein profile of goats infected by CAE and its associations with seroconversion using Western blotting. Two groups containing five males each, were used in this experiment. The first group was composed by seropositive animals and the control by seronegative confirmed by Western blotting and PCR. The semen was collected through artificial vagina and after that, two-dimensional electrophoresis and MALDI-TOF MS were used. Seventy-five spots were identified in the goat seminal plasma gels, equivalent to 13 different proteins with more expression. The similar proteins found in both groups and related to reproduction were spermadhesin Z13-like, bodhesin and bodhesin-2, Lipocalin, protein PDC-109-like, and albumin. In infected goats, proteases such as arisulfatase A have been identified, whose function probably is related to metabolism control of sulfatides, involved to virus control. The other ones were bifunctional ATP-dependent dihydroxyacetone kinase/FAD-AMP lyase, cathepsin F isoform X1, disintegrin and metalloproteinase domain-containing protein 2-like isoform X1, clusterin, carbonic anhydrase 2, electron transfer flavoprotein subunit beta, and epididymal secretory glutathione peroxidase. The results of this study show the reaction of the innate immune system against chronic infection of goats by CAE.


Assuntos
Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Doenças das Cabras/diagnóstico , Infecções por Lentivirus/veterinária , Proteínas de Plasma Seminal/análise , Animais , Western Blotting/veterinária , Eletroforese em Gel Bidimensional/veterinária , Doenças das Cabras/virologia , Cabras/genética , Infecções por Lentivirus/diagnóstico , Infecções por Lentivirus/virologia , Masculino , Reação em Cadeia da Polimerase/veterinária , Sêmen/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária
2.
J Vet Diagn Invest ; 30(1): 36-41, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28868985

RESUMO

Roughly one-fourth of goats infected with small ruminant lentivirus (SRLV) develop caprine arthritis-encephalitis (CAE). We compared the profile of antibody response to surface glycoprotein (SU), and combined transmembrane glycoprotein and capsid protein (TM/CA) in SRLV-infected arthritic and asymptomatic goats, and determined the ability of 2 commercial ELISAs to distinguish between arthritic and asymptomatic goats. We used sera from 312 SRLV-seropositive dairy goats in a whole-virus ELISA; 222 were collected from arthritic goats and 90 from apparently healthy goats. Sera were screened with a competitive inhibition ELISA based on SU antigen (SU-ELISA) and an indirect ELISA based on TM and CA antigens (TM/CA-ELISA). Receiver operating characteristic (ROC) curves were prepared for both ELISAs, and areas under the ROC curves (AUC) were compared. The proportion of goats with antibody response stronger to SU antigen than to TM/CA antigen was significantly higher among arthritic than asymptomatic goats (58.1% vs. 28.9%; p < 0.001). Antibody response to SU antigen was a good predictor of the arthritic form of CAE: AUC for SU-ELISA was 89.7% (95% CI: 85.2%, 94.2%), compared to 59.3% (95% CI: 51.9%, 66.8%) for TM/CA-ELISA ( p < 0.001). With the cutoff set at percentage of inhibition of 56%, SU-ELISA had sensitivity of 86.9% (95% CI: 81.9%, 90.7%) and specificity of 84.4% (95% CI: 75.6%, 90.5%) in discriminating between arthritic and asymptomatic goats.


Assuntos
Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças das Cabras/diagnóstico , Infecções por Lentivirus/veterinária , Glicoproteínas de Membrana/análise , Proteínas Virais/análise , Animais , Anticorpos Antivirais/sangue , Proteínas do Capsídeo/análise , Ensaios Enzimáticos/métodos , Ensaios Enzimáticos/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Doenças das Cabras/virologia , Cabras , Infecções por Lentivirus/diagnóstico , Infecções por Lentivirus/virologia , Polônia
3.
Vet Rec ; 182(2): 51, 2018 01 13.
Artigo em Inglês | MEDLINE | ID: mdl-29109181

RESUMO

Small ruminant lentivirus infections in goats affect both production and animal welfare. This represents a threat to the qualitative and quantitative growth of goat farming, recently observed in mountainous regions such as the Autonomous Province of Bolzano - South Tyrol (Italy). To monitor and eradicate the caprine arthritis encephalitis virus in this goat population, a compulsory eradication campaign was launched, based on a strict census of small ruminants and yearly serological testing of all animals, followed by the consequent culling of seropositive individuals. The campaign succeeded in completely eliminating cases of clinical disease in goats, while drastically reducing the seroprevalence at the herd as well as individual animal level. The serological outcome of the introduced control measures was determined using commercially available ELISA kits, demonstrating their suitability for use in this type of campaign, aimed at reducing seroprevalence as well as clinical manifestations of these infections. However, this clear success is diminished by the failure to achieve a complete eradication of these viruses. The reasons leading to the observed tailing phenomenon and the occurrence of new infections in already sanitised flocks are discussed and implementation of further measures are proposed.


Assuntos
Erradicação de Doenças , Doenças das Cabras/prevenção & controle , Doenças das Cabras/virologia , Infecções por Lentivirus/veterinária , Logro , Animais , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças das Cabras/epidemiologia , Cabras , Itália/epidemiologia , Infecções por Lentivirus/epidemiologia , Infecções por Lentivirus/prevenção & controle , Avaliação de Programas e Projetos de Saúde , Vigilância de Evento Sentinela/veterinária , Estudos Soroepidemiológicos , Testes Sorológicos/veterinária
4.
Arch Virol ; 162(10): 3007-3015, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28642977

RESUMO

Small ruminant lentiviruses (SRLVs), which comprise caprine arthritis-encephalitis virus (CAEV) and maedi-visna virus (MVV), are prevalent in goats and sheep worldwide, including in Japan. However, little is known about the molecular characteristics of goat lentiviruses in Japan. In this study, a molecular and phylogenetic analysis of the long gag region was performed. The phylogenic tree demonstrated that all samples belonged to SRLV subtype B1. Two clusters were identified, with one cluster distinct from previously reported strains of subtype B1. In addition, several alterations in the amino acid sequence were detected in immunodominant epitopes of the gag region. To gain a deeper understanding of the genetic diversity of SRLVs in Japan, it will be necessary to increase the sample size and conduct a broader survey. The present report is important for establishing baseline information on the prevalence of SRLV in Japan and providing data to develop a new, more sensitive diagnostic test for effective control of SRLV.


Assuntos
Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Doenças das Cabras/virologia , Cabras , Infecções por Lentivirus/veterinária , Vírus Visna-Maedi/isolamento & purificação , Visna/virologia , Animais , Feminino , Doenças das Cabras/epidemiologia , Japão , Infecções por Lentivirus/epidemiologia , Infecções por Lentivirus/virologia , Masculino , Ovinos , Visna/epidemiologia
5.
Pesqui. vet. bras ; 37(6): 577-581, jun. 2017. tab, graf
Artigo em Português | LILACS, VETINDEX | ID: biblio-895456

RESUMO

Objetivou-se no estudo identificar a ocorrência de infecção pelo vírus da artrite encefalite caprina (CAEV) em propriedades produtoras de leite caprino com sistema intensivo no estado de Minas Gerais. Foram avaliadas cinco propriedades, localizadas em cidades distintas, totalizando 1072 animais, sendo 48 machos e 1024 fêmeas de diferentes faixas etárias, das raças Toggenburg, Alpina e Saanen. O método de diagnóstico utilizado foi o de imunodifusão em ágar gel (IDGA), para detecção de anticorpos anti-CAEV, por ser o diagnóstico preconizado pela Organização Mundial de Saúde Animal (OIE). A ocorrência de anticorpos anti-CAEV nas propriedades estudadas foi de 49,5% (531/1072), entretanto a mesma variou de acordo com a propriedade. Foram observados os seguintes resultados por propriedade: propriedade 1 = 69,6% (156/224); propriedade 2 = 41,5% (47/113); propriedade 3 = 40,3% (63/156); propriedade 4 = 24,6% (18/73) e propriedade 5 = 48,8% (247/506). Do total de machos avaliados, 14,5% (7/48) apresentaram sorologia positiva. De acordo com os resultados, uma alta ocorrência de animais soropositivos foi identificada no estado de Minas Gerais, o qual possui um dos maiores rebanhos de cabras leiteiras do Brasil. Portanto, salienta-se a necessidade de se adotar medidas estratégias sanitárias para o controle da CAE, como a realização de exames rotineiros nas propriedades e a separação de animais infectados dos sadios. A exclusão de reprodutores positivos nas propriedades também é uma medida de controle, pois já foi demonstrado que estes são fontes de infecção importantes.(AU)


The objective of the present study was to identify the occurrence of caprine arthritis encephalitis virus (CAEV) infection in dairy goat farms with intensive system in Minas Gerais State. Five properties were evaluated, totaling 1072 animals, being 48 males and 1024 females of different ages and breeds (Toggenburg, Saanen and Alpine). The diagnostic method used was the agar gel immunodiffusion (AGID) test, to detect antibodies anti-CAEV, which is the diagnostic test recommended by World Organization for Animal Health - OIE. The occurrence of anti-CAEV antibodies in the properties was 49.5% (531/1072), however varied according to the farm. The anti-CAEV antibodies detection varied according to the property: Farm 1 = 69.6% (156/224); farm 2 = 41.5% (47/113); farm 3 = 40.3% (63/156); farm 4 = 24.6% (18/73) and farm 5 = 48.8% (247/506). Of the total sampled males, 14.5% (7/48) were serologically positive. According to the results, there is a high occurrence rate of CAEV-seropositive animals in Minas Gerais, which has one of the largest Brazilian dairy goat herds. Therefore, it is essential to adopt strategies for CAE control, such as a routine of diagnostics tests in the properties and the separation of healthy from infected goats. The elimination of positive breeding goats in the properties is also a measure of control, because it has been shown that these are an important route of CAEV transmission in dairy farms.(AU)


Assuntos
Animais , Cabras/virologia , Infecções por Lentivirus/epidemiologia , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Imunodifusão/veterinária , Gado/virologia
6.
Pesqui. vet. bras ; 37(6): 544-548, jun. 2017. tab, mapas
Artigo em Português | LILACS, VETINDEX | ID: biblio-895467

RESUMO

O objetivo deste estudo foi determinar a soroprevalência de lentivírus de pequenos ruminantes (LVPR) e identificar os fatores de risco para a ocorrência de caprinos e ovinos soropositivos no semiárido do Estado da Paraíba. Foram utilizados 1.733 animais, sendo 1.274 caprinos procedentes de 62 Unidades de Produção (UPs) e 459 ovinos provenientes de 32 UPs. Para o diagnóstico sorológico da infecção por lentivírus foi utilizado o teste de imunodifusão em gel de ágar (IDGA). Dos 1.274 caprinos analisados 15 (1,18%) foram soropositivos, enquanto que todos os 459 ovinos foram soronegativos. Das 62 propriedades caprinas analisadas oito (12,9%) apresentaram pelo menos um animal soropositivo. Os fatores de risco para a ocorrência de caprinos soropositivos foram área da propriedade (odds ratio = 3,28; p = 0,044), ausência de capacitação dos produtores (odds ratio = 8,29; p = 0,042) e uso de monta natural não controlada (odds ratio = 6,78; p = 0,012). Conclui-se que a infecção por lentivírus de pequenos ruminantes, demonstrada pela detecção de anticorpos, está disseminada em rebanhos caprinos do semiá­rido paraibano, e sugere-se o incentivo à capacitação contínua dos produtores, manutenção de reprodutores negativos ao LVPR e utilização de inseminação artificial com o intuito de evitar o contato físico entre macho e fêmeas.(AU)


The aim of this survey was to determine the seroprevalence of small ruminant lentivirus (SRLV) and to identify risk factors for the occurrence of seropositive goats and sheep in the semiarid region of Paraiba State. It were used 1,733 animals, being 1,274 goats from 62 Production Units (PU) and 459 sheep from 38 PU. For the serological diagnosis of lentivirus infection the agar gel immunodiffusion test (AGID) was used. Of the 1,274 goats 15 (1.18%) were seropositive, and all 459 sheep were seronegative. Of the 62 goat herds eight (12.9%) presented at least one seropositive animal. Risk factors for the occurrence of seropositive goats were area of the property ≤35 ha (odds ratio = 3.28; p=0.044), not training of producers (odds ratio = 8.29; p=0.042) and use of uncontrolled natural mating (odds ratio = 6.78; p=0.012). It is concluded that lentivirus infection detected by serology is spread in goat flocks in the semiarid of the State of Paraíba, and it is suggested to encourage the continous capacitation of owners, maintenance of reproducers negative for SRLV and use of artificial insemination aiming to avoid the physical contact among male and females.(AU)


Assuntos
Animais , Ruminantes , Ovinos , Infecções por Lentivirus/etiologia , Infecções por Lentivirus/epidemiologia , Vírus Visna-Maedi/isolamento & purificação , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Estudos Epidemiológicos , Fatores de Risco
7.
J Virol Methods ; 243: 98-104, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28159666

RESUMO

Caprine arthritis-encephalitis (CAE) in goats is a complex disease syndrome caused by a lentivirus. This persistent viral infection results in arthritis in adult goats and encephalitis in lambs. The prognosis for the encephalitic form is normally poor, and this form of the disease has caused substantial economic losses for goat farmers. Hence, a more efficient detection platform based on recombinase polymerase amplification (RPA) and a lateral flow dipstick (LFD) was developed in the present study for detecting the proviral DNA of caprine arthritis-encephalitis virus (CAEV). Under the optimal incubation conditions, specifically, 30min at 37°C for RPA followed by 5min at room temperature for LFD, the assay was found to be sensitive to a lower limit of 80pg of total DNA and 10 copies of plasmid DNA. Furthermore, there was no cross-reaction with other tested viruses, including goat pox virus and bovine leukemia virus. Given its simplicity and portability, this RPA-LFD protocol can serve as an alternative tool to ELISA for the primary screening of CAEV, one that is suitable for both laboratory and field application. When the RPA-LFD was applied in parallel with serological ELISA for the detection of CAEV in field samples, the RPA-LFD assay exhibited a higher sensitivity than the traditional method, and 82% of the 200 samples collected in Taiwan were found to be positive. To our knowledge, this is the first report providing evidence to support the use of an RPA-LFD assay as a specific and sensitive platform for detecting CAEV proviral DNA in goats in a faster manner, one that is also applicable for on-site utilization at farms and that should be useful in both eradication programs and epidemiological studies.


Assuntos
Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Cromatografia/métodos , Doenças das Cabras/diagnóstico , Doenças das Cabras/virologia , Infecções por Lentivirus/veterinária , Técnicas de Amplificação de Ácido Nucleico/métodos , Sistemas Automatizados de Assistência Junto ao Leito , Animais , Vírus da Artrite-Encefalite Caprina/genética , DNA Viral/genética , DNA Viral/isolamento & purificação , Cabras , Infecções por Lentivirus/diagnóstico , Provírus/genética , Recombinases/metabolismo , Sensibilidade e Especificidade , Taiwan , Temperatura
8.
Vet Rec ; 180(9): 226, 2017 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-28062845

RESUMO

The aim of the present transversal descriptive study was to determine the exposure and risk factors associated with caprine arthritis-encephalitis virus (CAEV) in southern Spain. A total of 3,312 serum samples were collected from goat belonging to three different breeds coming from 48 flocks located in different geographic areas from southern Spain. In addition, health and productive parameters were surveyed during the visit to the herds. Serum samples were analysed by INgezim Maedi Screening (Ingenasa®) ELISA kit. The total percentage of herds exposed to CAEV was 87.71% (CI95 78.42-97.00). A total of 733 goats were seropositive with overall seroprevalence of 23.22% (CI95 21.78-24.65). The intraherd seroprevalence was 20.82%±24.07. Multivariate logistic regression showed significant association between CAEV and the next variables: (i) herd size (P<0.0001; OR: 2.07; CI95: 1.73-2.50), (ii) kidding area (P<0.0020; OR: 1.38; CI95: 1.13-1.69), (iii) cleaning and disinfection program (P<0.0067; OR: 1.90; CI95: 1.43-2.53), (iv) natural mating (P<0.0026; OR: 2.22; CI95: 1.73-2.86) and (v) multiparous (P<0.0001; OR: 2.90; CI95: 2.17-3.87). The results indicate a widespread of CAEV infection in goat herds in southern Spain, being herd size, existence of kidding area, absence of cleaning and disinfection program, natural mating and multiparous goats risk factors for the exposure to CAEV.


Assuntos
Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Exposição Ambiental/estatística & dados numéricos , Doenças das Cabras/epidemiologia , Doenças das Cabras/virologia , Infecções por Lentivirus/veterinária , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Cabras , Infecções por Lentivirus/epidemiologia , Fatores de Risco , Estudos Soroepidemiológicos , Espanha/epidemiologia
9.
Trop Anim Health Prod ; 48(5): 1083-7, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26992736

RESUMO

This paper reports a first-time study performed in El Salvador on the presence or absence of antibodies to three important animal diseases in small ruminants. The work was conducted in the west and central departments of the country, selecting 42 and 43 cantons with an existing sheep and goat population, respectively. Serum samples were collected from 396 sheep and 335 goats and tested for seropositivity to Brucella (B.) spp. The specimens from goats were also tested for antibodies to caprine arthritis-encephalitis (CAE) virus. Four (1 %) sheep and none of the goats were seropositive by Rose Bengal test. All animals were negative by indirect ELISA (iELISA) for B. abortus. All animals were negative by iELISA for CAE. A total of 383 sheep and 330 goats underwent the single intradermal cervical tuberculin (SICT) test for tuberculosis. Seventy (18 %) sheep and 43 (13 %) goats reacted to the SICT test. Those reactors were subjected to the single intradermal comparative cervical tuberculin (SICCT) test, and one (0.3 %) goat was deemed to be a positive reactor. No mycobacteria were diagnosed in concluding analyses, and further studies are considered necessary to determine the prevalence of the investigated diseases. Additionally, it is recommended that small ruminants should be included in the national eradication program on bovine brucellosis and tuberculosis to prevent potential reservoirs.


Assuntos
Brucelose/veterinária , Doenças das Cabras/epidemiologia , Infecções por Lentivirus/veterinária , Doenças dos Ovinos/epidemiologia , Tuberculose/veterinária , Animais , Anticorpos Antibacterianos/análise , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Brucella/isolamento & purificação , Brucelose/epidemiologia , Brucelose/microbiologia , El Salvador/epidemiologia , Doenças das Cabras/microbiologia , Doenças das Cabras/virologia , Cabras , Infecções por Lentivirus/epidemiologia , Infecções por Lentivirus/virologia , Mycobacterium/isolamento & purificação , Prevalência , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/virologia , Carneiro Doméstico , Tuberculose/epidemiologia , Tuberculose/microbiologia
10.
Virology ; 487: 50-8, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26517396

RESUMO

Small ruminant lentiviruses infect goats and sheep, inducing clinical disease in a minority of infected animals. Following an eradication campaign, clinical cases may disappear in a population. The complete elimination of these lentiviruses is however difficult to achieve and the spreading of less virulent strains often parallels the elimination of their virulent counterparts. Here, we characterized three such strains isolated from a flock in the post-eradication phase. We completely sequenced their genomes, showing that one of the isolates was most probably the product of a recombination event between the other two viruses. By comparing the sequences of these isolates with those of virulent strains, we found evidence that particular LTR mutations may explain their attenuated phenotype. Finally, we constructed an infectious molecular clone representative of these viruses, analyzing its replication characteristics in different target cells. This clone will permit us to explore the molecular correlates of cytopathogenicity and virulence.


Assuntos
Vírus da Artrite-Encefalite Caprina/genética , Clonagem Molecular/métodos , Infecções por Lentivirus/virologia , RNA Viral/genética , Vírus Visna-Maedi/genética , Animais , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Vírus da Artrite-Encefalite Caprina/patogenicidade , Sequência de Bases , Células Cultivadas , Efeito Citopatogênico Viral/genética , Doenças das Cabras/virologia , Cabras , Macrófagos/virologia , Dados de Sequência Molecular , Alinhamento de Sequência , Análise de Sequência de RNA , Ovinos , Doenças dos Ovinos/virologia , Vírus Visna-Maedi/isolamento & purificação , Vírus Visna-Maedi/patogenicidade
11.
Arch Virol ; 160(4): 969-78, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25655265

RESUMO

Caprine arthritis encephalitis virus (CAEV) causes caprine arthritis encephalitis syndrome, which is an emerging disease of goats in the Philippines. DNA sequence analysis showed homology of 86-93 % between Philippine CAEV and available CAEV sequences in GenBank. CAEV was detected using nested polymerase chain reaction (PCR), and new sets of primers were designed in order to amplify the gag gene, which is a highly conserved region of the viral genome. In addition, the Philippine CAEV isolate clustered in group B with the prototype caprine lentivirus. Based on amino acid sequence alignments, it is possible that the Philippine CAEV isolate is a new strain of CAEV, but it is also possible that it was already present in the country even before the start of goat importation. Molecular characterization of the CAEV gag gene is important for the development of a detection kit specific for the local strain of CAEV and the establishment of small ruminant lentivirus eradication programs in the Philippines. This study is the first report to describe the molecular characteristics of CAEV circulating in the Philippines.


Assuntos
Vírus da Artrite-Encefalite Caprina/genética , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Produtos do Gene gag/genética , Doenças das Cabras/virologia , Infecções por Lentivirus/veterinária , Sequência de Aminoácidos , Animais , Vírus da Artrite-Encefalite Caprina/química , Vírus da Artrite-Encefalite Caprina/classificação , Produtos do Gene gag/química , Genoma Viral , Doenças das Cabras/epidemiologia , Cabras , Infecções por Lentivirus/epidemiologia , Infecções por Lentivirus/virologia , Dados de Sequência Molecular , Filipinas/epidemiologia , Filogenia , Alinhamento de Sequência
12.
Arq. bras. med. vet. zootec ; 67(1): 94-102, 2/2015. tab
Artigo em Português | LILACS | ID: lil-741121

RESUMO

Neste estudo, 67 ejaculados foram avaliados, antes e depois da técnica de swim-up, em relação à qualidade seminal e à presença do CAEV. Das 67 amostras testadas por PCRn, antes do swim-up, 47 (70,15%) foram positivas para o DNA pró-viral. No entanto, quatro amostras adicionais foram positivas ao RT-nested PCR após o swim-up, o que permite dizer que, pelo menos, 76,12% (51/67) delas estavam infectadas antes da lavagem. Todavia, em 23,88% (16/67) das amostras não foi detectada a presença do CAEV. Após a aplicação da técnica de swim-up, constatou-se, pela PCRn e RT-nested PCR, que houve uma redução significativa (χ²= 9,078; p<0,001) da presença do CAEV nas amostras seminais, pois 28 de 51 amostras positivas resultaram livres do vírus (54,90%), tanto para DNA pró-viral quanto para o vírus livre. Em relação à motilidade individual progressiva (MIP) e vigor espermático obtidos antes e depois da técnica de swim-up, observou-se uma diminuição significativa em suas médias, sendo o MIP de 86,42% para 71,49%, já o vigor espermático de 4,16 para 3,93. Conclui-se que a eliminação do CAEV no sêmen é de caráter intermitente, e que a associação da PCRn e RT-nested PCR é uma opção segura para a certificação sanitária individual das amostras seminais quanto à presença ou ausência do CAEV. Finalmente, a técnica de swim-up promove uma redução na infectividade de amostras de sêmen contaminadas, e, além disso, é possível promover a recuperação de espermatozoides viáveis.


In this study, 67 ejaculates were assessed before and after the swim-up technique in relation to semen quality and presence of CAEV. Of the 67 samples tested by Nested PCR, before swim-up 47 (70.15%) were positive for viral DNA. Furthermore, four additional samples were positive for RT-nested PCR after swim-up, which allows us to affirm that at least 76.12% (51/67) were infected before washing. However, 23.88% (16/67) of the samples did not detect the presence of CAEV. After application of the swim-up technique it was found, by Nested PCR and RT-nested PCR, that there was a significant decrease (χ² = 9.078, p <0.001) in the presence of CAEV in semen samples, once 28 of 51 positive samples were free from the virus (54.90%) for both proviral DNA and the free form of the virus. Regarding individual progressive motility (IPM) and spermatic vigor obtained before and after the swim-up technique, a significant decrease was observed in the average, being 86.42% of the IPM to 71.49% and the spermatic vigor from 4.16 for the 3.93. It is concluded that the removal of CAEV in semen has an intermittent character, and the combination of PCR and RT-nested PCR is a safe option for health certification of individual semen samples for the presence or absence of CAEV. Finally, the swim-up technique promotes a reduction in the infectivity of contaminated semen samples, and it is possible to promote the recovery of high individual progressive motility sperm and sperm vigor.


Assuntos
Animais , Masculino , Reação em Cadeia da Polimerase/estatística & dados numéricos , Reação em Cadeia da Polimerase/veterinária , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Análise do Sêmen/veterinária
13.
Rev Sci Tech ; 34(3): 915-21, 907-14, 2015 Dec.
Artigo em Inglês, Francês | MEDLINE | ID: mdl-27044161

RESUMO

An epidemiological survey, accompanied by a serological analysis,was conducted on samples taken from Lebanese goat herds in order to determine the prevalence of infection with the caprine arthritis encephalitis virus (CAEV) in Lebanon. The results of the survey provided information on various livestock production, animal health and herd management factors. Serum samplesfrom 952 goats, including the local breeds (Baladi and Damascene) and imported breeds (Alpine and Saneen), were taken from 60 farms distributed throughout Lebanon and tested for the presence of anti-CAEV antibodies. The data obtained were analysed using a statistical model to assess CAEV infection risk factors in Lebanon. In total, 125 samples proved to be positive, representing a prevalence in selected individuals of 13.1% and in selected herds of 51.7%. The Bekaa region had the highest number of herds with seropositive goats (90% of herds); the level was lower in Mount Lebanon, the North and the South (54%, 34% and 33%, respectively). The prevalence in relation to the livestock production system was 70% in herds in intensive systems, 54% in semi-intensive systems and 45% in extensive systems. The indigenous breeds were more resistant and tolerant of CAEV than the imported breeds. This study confirms the presence of CAEV in Lebanese goat herds and identifies the different livestock production practices likely to favour the rapid spread of the virus.


Assuntos
Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Doenças das Cabras/virologia , Infecções por Lentivirus/veterinária , Animais , Doenças das Cabras/epidemiologia , Cabras , Líbano/epidemiologia , Infecções por Lentivirus/epidemiologia , Infecções por Lentivirus/virologia , Fatores de Risco
14.
Vet Rec ; 176(7): 173, 2015 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-25344573

RESUMO

The objective of this study was to evaluate the diagnostic performance of two ELISA tests applied to bulk tank milk (BTM) as the first part of a two-step test scheme for the surveillance of caprine arthritis encephalitis (CAE) and caseous lymphadenitis (CLA) infections in goats. The herd-level BTM tests were assessed by comparing them to the test results of individual serological samples. The potential for refining the cut-off levels for BTM tests used as surveillance tools in a population recently cleared of infection was also investigated. Data was gathered on serum (nCAE =9702 and nCLA=13426) and corresponding BTM (nCAE=78 and nCLA=123) samples from dairy goat herds enrolled in the Norwegian disease control and eradication programme 'Healthier Goats'. The results showed that the sensitivity and specificity of the CAE ELISA BTM test with respect to detecting ≥2 per cent within-herd prevalence were 72.7 per cent and 86.6 per cent, respectively. For the CLA ELISA BTM the sensitivity and specificity were 41.4 per cent and 81.7 per cent, respectively, for the same goal of detection. The results suggest that BTM testing can be applied as a cost-effective first step for early detection of CAE and CLA infection.


Assuntos
Infecções por Corynebacterium/veterinária , Doenças das Cabras/diagnóstico , Infecções por Lentivirus/veterinária , Linfadenite/veterinária , Leite/microbiologia , Animais , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Infecções por Corynebacterium/sangue , Infecções por Corynebacterium/diagnóstico , Infecções por Corynebacterium/epidemiologia , Corynebacterium pseudotuberculosis/isolamento & purificação , Análise Custo-Benefício , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Doenças das Cabras/sangue , Doenças das Cabras/epidemiologia , Cabras , Infecções por Lentivirus/sangue , Infecções por Lentivirus/diagnóstico , Infecções por Lentivirus/epidemiologia , Linfadenite/sangue , Linfadenite/diagnóstico , Linfadenite/epidemiologia , Leite/virologia , Noruega/epidemiologia , Vigilância da População/métodos , Sensibilidade e Especificidade , Testes Sorológicos/veterinária
15.
Virol J ; 11: 65, 2014 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-24708706

RESUMO

BACKGROUND: Small ruminant lentiviruses escaping efficient serological detection are still circulating in Swiss goats in spite of a long eradication campaign that essentially eliminated clinical cases of caprine arthritis encephalitis in the country. This strongly suggests that the circulating viruses are avirulent for goats.To test this hypothesis, we isolated circulating viruses from naturally infected animals and tested the in vitro and in vivo characteristics of these field isolates. METHODS: Viruses were isolated from primary macrophage cultures. The presence of lentiviruses in the culture supernatants was monitored by reverse transcriptase assay. Isolates were passaged in different cells and their cytopathogenic effects monitored by microscopy. Proviral load was quantified by real-time PCR using customized primer and probes. Statistical analysis comprised Analysis of Variance and Bonferroni Multiple Comparison Test. RESULTS: The isolated viruses belonged to the small ruminant lentiviruses A4 subtype that appears to be prominent in Switzerland. The 4 isolates replicated very efficiently in macrophages, displaying heterogeneous phenotypes, with two isolates showing a pronounced cytopathogenicity for these cells. By contrast, all 4 isolates had a poor replication capacity in goat and sheep fibroblasts. The proviral loads in the peripheral blood and, in particular, in the mammary gland were surprisingly high compared to previous observations. Nevertheless, these viruses appear to be of low virulence for goats except for the mammary gland were histopathological changes were observed. CONCLUSIONS: Small ruminant lentiviruses continue to circulate in Switzerland despite a long and expensive caprine arthritis encephalitis virus eradication campaign. We isolated 4 of these lentiviruses and confirmed their phylogenetic association with the prominent A4 subtype. The pathological and histopathological analysis of the infected animals supported the hypothesis that these A4 viruses are of low pathogenicity for goats, with, however, a caveat about the potentially detrimental effects on the mammary gland. Moreover, the high proviral load detected indicates that the immune system of the animals cannot control the infection and this, combined with the phenotypic plasticity observed in vitro, strongly argues in favour of a continuous and precise monitoring of these SRLV to avoid the risk of jeopardizing a long eradication campaign.


Assuntos
Vírus da Artrite-Encefalite Caprina/genética , Vírus da Artrite-Encefalite Caprina/patogenicidade , Doenças das Cabras/virologia , Infecções por Lentivirus/veterinária , Animais , Vírus da Artrite-Encefalite Caprina/classificação , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Sangue/virologia , Células Cultivadas , Análise por Conglomerados , Efeito Citopatogênico Viral , Fibroblastos/virologia , Genótipo , Doenças das Cabras/epidemiologia , Cabras , Humanos , Infecções por Lentivirus/epidemiologia , Infecções por Lentivirus/virologia , Macrófagos/virologia , Glândulas Mamárias Humanas/virologia , Microscopia , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Provírus/genética , Provírus/isolamento & purificação , RNA Viral/genética , Análise de Sequência de DNA , Ovinos , Suíça/epidemiologia , Carga Viral
16.
Arq. bras. med. vet. zootec ; 66(2): 417-424, Jan.-Apr. 2014. ilus, tab
Artigo em Português | LILACS | ID: lil-709279

RESUMO

A artrite-encefalite caprina (CAE) é diagnosticada rotineiramente pela técnica de imunodifusão em gel de agarose (IDGA), que é considerada pouco sensível. Objetivou-se com este estudo padronizar testes de Elisa-i e Western Blot (WB) para diagnóstico precoce de anticorpos em caprinos contra CAEV e comparar os resultados obtidos nesses testes com a prova de IDGA. Para a padronização dos testes Elisa-i e WB, utilizaram-se diferentes concentrações e diluições de antígeno, soros e conjugado. No Elisa-i, adotaram-se microplacas rígidas com 96 poços, sendo a combinação de concentração de 0,5µg/poço de antígeno e diluições de 1:100 de soro e 1:1500 de conjugado a que apresentou melhor resultado. No WB foram utilizadas membranas de nitrocelulose, definindo-se as diluições de 1:50 de soro e 1:15000 de conjugado. Para avaliar o desempenho das técnicas, 222 amostras de soro caprino foram testadas e os dados obtidos foram comparados com o IDGA. A sensibilidade e a especificidade do Elisa-i/IDGA, WB/IDGA e WB/Elisa-i foram de 70% e 91%, 100% e 72,6%, 84,6% e 76,5%, concomitantemente. O índice Kappa desses testes foi de 0,35, 0,2 e 0,36, respectivamente. As técnicas de Elisa-i e WB apresentaram-se mais sensíveis que a IDGA, podendo ser utilizadas como ferramentas para o diagnóstico precoce da CAE...


Caprine arthritis-encephalitis (CAE) is routinely diagnosed with the Agarose Gel Immunodiffusion (AGID) technique, which is considered to have low sensitivity. The objective of this study was to standardize testing i-Elisa and Western Blot for early detection of antibodies against CAEV in goats and compare the results obtained in these tests with proof of AGID. For standardization of i-Elisa and WB, different concentrations and dilutions of antigen, sera and conjugate were used. In the i-Elisa, rigid microplate with 96 wells was adopted, and the combination that showed the best result was a concentration of 0.5µg/ well of antigen and dilutions of the serum of 1:100 and conjugate of 1:1500. In the WB nitrocellulose membranes were used, and the dilutions of the serum were defined at 1:50 and conjugate at 1:15000. To evaluate the performance of the techniques, 222 goat serum samples were tested and the data were compared with the AGID. The sensitivity and specificity of Elisa-i/IDGA, WB/AGID and WB/Elisa-i were 70% and 91%, 100% and 72.6%, 84.6% and 76.5%, concomitantly. The Kappa index of these tests was 0.35, 0.2 and 0.36, respectively. The i-Elisa and WB techniques were more sensitive than the AGID and can be used as tools for early diagnosis of CAE...


Assuntos
Animais , Cabras/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Imunodifusão/veterinária , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Western Blotting/veterinária
17.
Diagn Microbiol Infect Dis ; 79(1): 37-42, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24630755

RESUMO

Caprine arthritis encephalitis virus (CAEV), of the genus Lentivirus of the Retroviridae family, causes persistent disease, which is characterized by polyarthritis and mastitis in adult goats and progressive paresis (leukoencephalomyelitis) in kids. A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of CAEV in blood samples. Species-specific primers amplifying the gag gene region in the provirus were used for the detection of CAEV. The LAMP assay result was obtained 30 min after incubation on a constant temperature at 63 °C in a heat block. Resulting amplicons were visualized by addition of SYBR green dye after the reaction and checked by agarose gel electrophoresis. The sensitivity of LAMP assay was evaluated by comparing the result with the nested polymerase chain reaction. Based on the experiments, the result of the assay indicated a rapid and sensitive test for the detection of CAEV.


Assuntos
Vírus da Artrite-Encefalite Caprina/genética , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Produtos do Gene gag/genética , Infecções por Lentivirus/virologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Animais , Eletroforese em Gel de Ágar , Cabras , Infecções por Lentivirus/diagnóstico
18.
Viruses ; 5(10): 2614-23, 2013 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-24153063

RESUMO

The diagnosis of Small Ruminant Lentivirus (SRLV) is based on clinical signs, pathological lesions and laboratory testing. No standard reference test for the diagnosis of maedi visna has been validated up to the present, and it is puzzling that tests which detect antibodies against the virus and tests which detect the proviral genome may render opposite results. The aim of this study was to evaluate the presence in milk throughout a lactation period of specific antibodies by ELISA and of SRLV proviral DNA by a PCR of the highly conserved pol region. A six-month study was conducted with the milk of 28 ewes and 31 goats intensively reared. The percentage of animals with antibodies against SRLV increased throughout the study period. Seroprevalence in sheep was 28% at the beginning of the study and by the end it had increased up to 52.4%. In goats, initial seroprevalence of 5.6% increased to 16%. The percentage of PCR positive ewes was stable throughout the study period. Of the positive sheep, 21.4% were PCR-positive before antibodies could be detected and most of them became PCR-negative shortly after the first detection of antibodies. This might suggest that antibodies have a neutralizing effect. In addition, an equal percentage of sheep were always PCR-negative but either became ELISA-positive or was always ELISA-positive, which might support this hypothesis. On the other hand, the PCR results in goats did not follow any pattern and oscillated between 35.3% and 55.6% depending on the month. Most goats positive by PCR failed to develop antibodies in the 6 months tested. We may conclude that the infection and the antibody response to it follow a different trend in sheep and goats.


Assuntos
Anticorpos/análise , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , DNA Viral/isolamento & purificação , Infecções por Lentivirus/veterinária , Leite/imunologia , Leite/virologia , Provírus/isolamento & purificação , Animais , Vírus da Artrite-Encefalite Caprina/genética , DNA Viral/genética , Ensaio de Imunoadsorção Enzimática , Doenças das Cabras/imunologia , Doenças das Cabras/virologia , Cabras , Infecções por Lentivirus/imunologia , Infecções por Lentivirus/virologia , Reação em Cadeia da Polimerase , Provírus/genética , Ovinos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/virologia , Fatores de Tempo
19.
Can Vet J ; 54(10): 960-4, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24155416

RESUMO

Computed tomography was used to aid in the antemortem diagnosis of leukoencephalomyelitis in a goat infected by caprine arthritis encephalitis virus (CAEV). Imaging results were corroborated by histologic examination. This report discusses various methods of imaging the nervous system and their potential for use in the antemortem diagnosis of CAEV neurologic changes.


Assuntos
Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Doenças das Cabras/virologia , Leucoencefalopatias/veterinária , Tomografia Computadorizada por Raios X/veterinária , Animais , Feminino , Doenças das Cabras/patologia , Cabras , Leucoencefalopatias/virologia
20.
Viruses ; 5(8): 2005-18, 2013 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-23955501

RESUMO

Caprine arthritis encephalitis virus (CAEV) is a lentivirus that infects both goats and sheep and is closely related to maedi-visna virus that infects sheep; collectively, these viruses are known as small ruminant lentiviruses (SRLV). Infection of goats and sheep with SRLV typically results in discrete inflammatory diseases which include arthritis, mastitis, pneumonia or encephalomyelitis. SRLV-infected animals concurrently demonstrating lentivirus-associated lesions in tissues of lung, mammary gland, joint synovium and the central nervous system are either very rare or have not been reported. Here we describe a novel CAEV promoter isolated from a sheep with multisystemic lentivirus-associated inflammatory disease including interstitial pneumonia, mastitis, polyarthritis and leukomyelitis. A single, novel SRLV promoter was cloned and sequenced from five different anatomical locations (brain stem, spinal cord, lung, mammary gland and carpal joint synovium), all of which demonstrated lesions characteristic of lentivirus associated inflammation. This SRLV promoter isolate was found to be closely related to CAEV promoters isolated from goats in northern California and other parts of the world. The promoter was denoted CAEV-ovine-MS (multisystemic disease); the stability of the transcription factor binding sites within the U3 promoter sequence are discussed.


Assuntos
Estruturas Animais/virologia , Vírus da Artrite-Encefalite Caprina/isolamento & purificação , Infecções por Lentivirus/veterinária , Regiões Promotoras Genéticas , Doenças dos Ovinos/virologia , Tropismo Viral , Animais , Artrite/veterinária , Artrite/virologia , Vírus da Artrite-Encefalite Caprina/genética , Vírus da Artrite-Encefalite Caprina/fisiologia , California , Infecções por Lentivirus/virologia , Doenças Pulmonares Intersticiais/veterinária , Doenças Pulmonares Intersticiais/virologia , Mastite/veterinária , Mastite/virologia , Dados de Sequência Molecular , RNA Viral/genética , Análise de Sequência de DNA , Ovinos
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