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1.
Rev. argent. microbiol ; 51(2): 119-129, jun. 2019. ilus, tab
Artigo em Inglês | LILACS | ID: biblio-1013360

RESUMO

Equid alphaherpesvirus 1 (EHV-1) infection causes abortion, respiratory disease, perinatal deaths and neurological disorders in horses. The natural infection and available vaccines provide only partial and short-lived protection against reinfections. In the present study, we analyzed the ability of purified baculovirus-expressed glycoprotein D (gD) administered by different routes to induce protective immunity in BALB/c mice after challenge with the EHV-1 AR8 strain. Clinical signs varied among the different groups of mice immunized by parenteral routes, and, although gD induced a specific serum IgG response, it did not prevent the virus from reaching the lungs. Intranasally immunized mice showed no clinical signs, and virus isolation from lungs, histological lesions and antigen detection by immunohistochemistry were negative. In addition, by this route, gD did not stimulate the production of serum IgG and IgA. However, a specific IgA response in the respiratory tract was confirmed in intranasally immunized mice. Thus, we conclude that the mucosal immune response could reduce the initial viral attachment and prevent the virus from reaching the lungs. Our findings provide additional data to further study new immunization strategies in the natural host.


La infección con alfaherpesvirus equino 1 (EHV-1) causa abortos, enfermedad respiratoria, muertes perinatales y desórdenes neurológicos en equinos. La infección natural y las vacunas disponibles solo proporcionan protección parcial y de corta duración contra las reinfecciones. En el presente estudio se analizó la inducción de inmunidad protectiva de la glicoproteina D (gD) expresada en baculovirus y purificada al ser administrada por diferentes rutas en ratones BALB/c desafiados con la cepa AR8 de EHV-1. Los signos clínicos fueron variables entre los grupos de ratones inmunizados por rutas parenterales y, aunque la gD indujo respuesta especifica de IgG en suero, no logró prevenir la llegada del virus al pulmón. En los ratones inmunizados intranasalmente no se observaron signos clinicos ni lesiones histopatológi-cas, y el aislamiento viral y la detección de antigenos por inmunohistoquímica en pulmón fueron negativos. Además, por esta ruta la gD no estimuló la producción de IgG y de IgA en suero. Sin embargo se confirmó la respuesta de IgA especifica en el tracto respiratorio de ratones inmunizados intranasalmente. Esta respuesta inmune mucosal podría haber reducido la unión inicial del virus a la célula huésped y, de este modo, prevenir la llegada del virus al pulmón. Nuestros hallazgos proporcionan un aporte para continuar estudiando nuevas estrategias de inmunización en el huésped natural.


Assuntos
Doenças Respiratórias/imunologia , Glicoproteínas/imunologia , Herpesvirus Equídeo 1/patogenicidade , Imuno-Histoquímica/veterinária , Imunização/veterinária , Cavalos/imunologia , Imunidade/efeitos dos fármacos
2.
Open Vet J ; 9(1): 33-37, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31086763

RESUMO

Bovine leukemia virus (BLV) is the agent responsible for enzootic bovine leukosis, the most common neoplastic disease in cattle. The horn fly, a major hematophagous pest of cattle, is able to transmit different diseases in cattle. However, its implication in BLV transmission under a natural environment is still discussed. The objectives of this work were to determine the presence of BLV in horn flies (by sequencing) and to evaluate the ability of horn flies to transmit BLV to cattle (through an experimental assay under a natural environment). To demonstrate the presence of BLV in the flies, 40 horn flies were collected from a BLV-positive cow with a sweep net and 10 pools with four horn-fly mouthparts each were prepared. The presence of BLV was determined by nested polymerase chain reaction and sequencing. To demonstrate BLV transmission, other 40 flies were collected from the same BLV-positive cow with a sweep net. Eight homogenates containing five horn-fly mouthparts each were prepared and injected to eight cows of different breeds, and blood samples were collected every 21 days. Then, to evaluate the ability of horn flies to transmit BLV to grazing cattle under natural conditions, both infected and uninfected cattle from the experimental transmission assay were kept together in the same paddock with more than 200 horn flies per animal for 120 days. Blood samples were collected every 20 days and the number of flies was determined. The sequencing results confirmed the presence of the provirus in horn flies. The results also confirmed that BLV transmission is a possible event, at least experimentally. However, the role of horn flies as vectors of BLV under a natural grazing system is still discussed.


Assuntos
Leucose Enzoótica Bovina/transmissão , Insetos Vetores/virologia , Vírus da Leucemia Bovina/isolamento & purificação , Muscidae/virologia , Animais , Argentina , Bovinos , Feminino , Insetos Vetores/fisiologia , Muscidae/fisiologia , Reação em Cadeia da Polimerase/veterinária , Provírus/isolamento & purificação
3.
Rev Argent Microbiol ; 51(2): 119-129, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30385072

RESUMO

Equid alphaherpesvirus 1 (EHV-1) infection causes abortion, respiratory disease, perinatal deaths and neurological disorders in horses. The natural infection and available vaccines provide only partial and short-lived protection against reinfections. In the present study, we analyzed the ability of purified baculovirus-expressed glycoprotein D (gD) administered by different routes to induce protective immunity in BALB/c mice after challenge with the EHV-1 AR8 strain. Clinical signs varied among the different groups of mice immunized by parenteral routes, and, although gD induced a specific serum IgG response, it did not prevent the virus from reaching the lungs. Intranasally immunized mice showed no clinical signs, and virus isolation from lungs, histological lesions and antigen detection by immunohistochemistry were negative. In addition, by this route, gD did not stimulate the production of serum IgG and IgA. However, a specific IgA response in the respiratory tract was confirmed in intranasally immunized mice. Thus, we conclude that the mucosal immune response could reduce the initial viral attachment and prevent the virus from reaching the lungs. Our findings provide additional data to further study new immunization strategies in the natural host.


Assuntos
Infecções por Herpesviridae/prevenção & controle , Herpesvirus Equídeo 1 , Proteínas do Envelope Viral/uso terapêutico , Animais , Modelos Animais de Doenças , Imunização , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/uso terapêutico , Proteínas do Envelope Viral/imunologia
4.
Arq. Inst. Biol ; (85): 1-7, Out 25, 2018.
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IPPROD, Sec. Est. Saúde SP | ID: biblio-1059868

RESUMO

Equine herpesvirus type 1 (EHV-1) is an important pathogen that causes abortion, neonatal disease, respiratory disorders, and neurological syndrome in equine populations worldwide. To evaluate EHV-1 as a cause of abortion and perinatal mortality in Brazil, tissue samples from 105 aborted equine fetuses, stillbirths, and foals up to one month of age were examined using virus isolation, immunohistochemistry (IHC), histopathology, and nested polymerase chain reaction (PCR). Two fetuses were positive for EHV-1 by PCR, one of which showed syncytia and eosinophilic intranuclear inclusion bodies in bronchial epithelia, but it was negative by virus isolation. The other showed no characteristic histological lesions, but it was positive by viral isolation. No sample was positive by IHC. The results presented low occurrence of EHV-1 in the studied population and suggested that the use of a combination of techniques increases the likelihood of an accurate diagnosis of EHV-1.


O herpes-vírus equino tipo 1 (HVE-1) é um importante agente patogênico causador de aborto, doença neonatal, distúrbios respiratórios e síndrome neurológica em populações de equinos em todo o mundo. Para avaliar a ocorrência do HVE-1 como agente causal de abortamento e mortalidade perinatal no Brasil, foram examinadas amostras de 105 fetos equinos abortados, natimortos e potros de até 1 mês de idade, utilizando as técnicas de isolamento viral, imuno-histoquímica (IHQ), histopatologia e reação em cadeia da polimerase aninhada (nested-PCR). Dois fetos foram positivos na análise de PCR, e um deles apresentou corpúsculos de inclusão viral eosinofílicos e sincícios no epitélio brônquico, porém foi negativo na análise de isolamento viral. O outro feto não apresentou lesões histológicas características de infecção herpética, mas foi positivo na análise de isolamento viral. Nenhuma amostra apresentou resultado positivo pela análise de IHQ. Os resultados demonstraram baixa ocorrência de HVE-1 na população estudada e que o uso de diferentes técnicas diagnósticas aumenta a probabilidade de um diagnóstico preciso para o HVE-1.


Assuntos
Herpesvirus Equídeo 1 , Imuno-Histoquímica , Reação em Cadeia da Polimerase , Vírus/isolamento & purificação
5.
Arq. Inst. Biol ; 85: e0852017, 2018. ilus, tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-998428

RESUMO

Equine herpesvirus type 1 (EHV-1) is an important pathogen that causes abortion, neonatal disease, respiratory disorders, and neurological syndrome in equine populations worldwide. To evaluate EHV-1 as a cause of abortion and perinatal mortality in Brazil, tissue samples from 105 aborted equine fetuses, stillbirths, and foals up to one month of age were examined using virus isolation, immunohistochemistry (IHC), histopathology, and nested polymerase chain reaction (PCR). Two fetuses were positive for EHV-1 by PCR, one of which showed syncytia and eosinophilic intranuclear inclusion bodies in bronchial epithelia, but it was negative by virus isolation. The other showed no characteristic histological lesions, but it was positive by viral isolation. No sample was positive by IHC. The results presented low occurrence of EHV-1 in the studied population and suggested that the use of a combination of techniques increases the likelihood of an accurate diagnosis of EHV-1.(AU)


O herpes-vírus equino tipo 1 (HVE-1) é um importante agente patogênico causador de aborto, doença neonatal, distúrbios respiratórios e síndrome neurológica em populações de equinos em todo o mundo. Para avaliar a ocorrência do HVE-1 como agente causal de abortamento e mortalidade perinatal no Brasil, foram examinadas amostras de 105 fetos equinos abortados, natimortos e potros de até 1 mês de idade, utilizando as técnicas de isolamento viral, imuno-histoquímica (IHQ), histopatologia e reação em cadeia da polimerase aninhada (nested-PCR). Dois fetos foram positivos na análise de PCR, e um deles apresentou corpúsculos de inclusão viral eosinofílicos e sincícios no epitélio brônquico, porém foi negativo na análise de isolamento viral. O outro feto não apresentou lesões histológicas características de infecção herpética, mas foi positivo na análise de isolamento viral. Nenhuma amostra apresentou resultado positivo pela análise de IHQ. Os resultados demonstraram baixa ocorrência de HVE-1 na população estudada e que o uso de diferentes técnicas diagnósticas aumenta a probabilidade de um diagnóstico preciso para o HVE-1.(AU)


Assuntos
Animais , Herpesvirus Equídeo 1/patogenicidade , Cavalos , Imuno-Histoquímica/métodos , Reação em Cadeia da Polimerase/métodos , Aborto Animal
6.
Rev Argent Microbiol ; 49(3): 210-215, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28551308

RESUMO

In this study we developed an indirect ELISA to detect antibodies against Minute Virus of Mice (MVM) using an antigen produced from BHK-21 cells infected with a prototype strain of the virus. The optimal antigen concentration and serum dilutions were established. In order to analyze variability in the laboratory, reproducibility and repeatability within and between plates were determined. Then, a panel of 460 sera from conventional facilities and previously classified as positive or negative by the indirect fluorescent antibody assay was analyzed. The cutoff value was determined by a receiver operating characteristic (ROC) curve. The results of the indirect ELISA were compared with those of the indirect fluorescent antibody assay. The ELISA assay showed 100% sensitivity and 99% specificity. ELISA is a useful tool to be developed in standard virology laboratories and can be used for screening animals faster than the traditional indirect fluorescent antibody assay.


Assuntos
Anticorpos Antivirais , Ensaio de Imunoadsorção Enzimática , Vírus Miúdo do Camundongo , Animais , Anticorpos Antivirais/análise , Técnica Indireta de Fluorescência para Anticorpo , Camundongos , Vírus Miúdo do Camundongo/imunologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
7.
Open Vet J ; 7(1): 32-35, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28331831

RESUMO

Caprine arthritis encephalitis virus (CAEV) has been reported in different countries worldwide, based on serological and molecular detection. In Argentina, the prevalence of CAEV infections is increasing, with goats showing symptoms associated mostly with cachexia and arthritis. Although in Argentina the virus has been detected by serology, it has never been isolated or characterized. Thus, the objectives of this work were to isolate and analyze the nucleotide sequences of the gag gene of Argentine CAEV strains and compare them with those of other SRLVs previously reported. Nucleotide sequence comparison showed homology with CAEV-Co, the CAEV prototype. Phylogenetic analyses showed that the Argentine strains clustered with genotype B, subtype B1. Because the molecular characterization of the gag region is suitable for phylogenetic studies and may be applied to monitor the control of SRLV, molecularly characterizing the Argentine CAEV strains may help develop a proper plan of eradication of CAEV infections.

9.
J Virol Methods ; 202: 15-8, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24631347

RESUMO

Equine herpesvirus 1 and 4 (EHV-1 and 4) infect most of the world's horses, causing serious clinical illness. Viral glycoproteins have been identified as the immunodominant antigens that generate the antiviral serological responses to EHV-1 and EHV-4 in infected horses. Here, glycoprotein D of EHV-1 was expressed by a recombinant baculovirus, purified and evaluated by a simple agar gel immunodiffusion test (AGID). Compared with virus neutralization, serological analysis by AGID showed good specificity (100%) and sensitivity (99.5%). The estimated Kappa values for repeatability and reproducibility were satisfactory. Thus, this rapid, inexpensive, simple and highly specific AGID test seems to be a valuable alternative tool for serological detection of antibodies against both EHV-1 and EHV-4.


Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/isolamento & purificação , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/virologia , Imunodifusão/métodos , Medicina Veterinária/métodos , Proteínas Virais , Animais , Glicoproteínas/genética , Glicoproteínas/isolamento & purificação , Infecções por Herpesviridae/diagnóstico , Infecções por Herpesviridae/virologia , Cavalos , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Testes Sorológicos/métodos , Proteínas Virais/genética , Proteínas Virais/isolamento & purificação
10.
Arch Virol ; 159(1): 141-5, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23851654

RESUMO

Here, we used a murine model to describe and compare the pathogenic potential of the Argentinean equid herpesvirus 1 (EHV-1) AR8 strain with the Japanese HH1 reference strain. In AR8-inoculated animals, clinical signs began earlier, but the viremic phase was shorter. Virus isolation and DNA detection in the lungs, liver and spleen were positive for both strains at different times postinfection (pi). Infection foci produced by both strains were immunohistochemically detected in lungs from day 1 to day 4 pi. We conclude that whichever EHV-1 strain is selected to experimentally reproduce the disease, it needs appropriate standardization in order to provide valid conclusions.


Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/fisiologia , Doenças dos Cavalos/virologia , Animais , Modelos Animais de Doenças , Infecções por Herpesviridae/patologia , Infecções por Herpesviridae/virologia , Herpesvirus Equídeo 1/classificação , Herpesvirus Equídeo 1/genética , Doenças dos Cavalos/patologia , Cavalos , Masculino , Camundongos , Camundongos Endogâmicos BALB C
11.
Rev. argent. microbiol ; 45(4): 222-228, Dec. 2013. ilus, tab
Artigo em Inglês | BINACIS | ID: bin-130223

RESUMO

Equine influenza virus is a leading cause of respiratory disease in horses worldwide. Disease prevention is by vaccination with inactivated whole virus vaccines. Most current influenza vaccines are generated in embryonated hens eggs. Virions are harvested from allantoic fluid and chemically inactivated. Although this system has served well over the years, the use of eggs as the substrate for vaccine production has several well-recognized disadvantages (cost, egg supply, waste disposal and yield in eggs). The aim of this study was to evaluate a baculovirus system as a potential method for producing recombinant equine influenza hemagglutinin to be used as a vaccine. The hemagglutinin ectodomain (HA1 subunit) was cloned and expressed using a baculovirus expression vector. The expression was determined by SDS-PAGE and immunoblotting. A high yield, 20 μg/ml of viral protein, was obtained from recombinant baculovirus-infected cells. The immune response in BALB/c mice was examined following rHA1 inoculation. Preliminary results show that recombinant hemagglutinin expressed from baculovirus elicits a strong antibody response in mice; therefore it could be used as an antigen for subunit vaccines and diagnostic tests.(AU)


El virus de la influenza equina es una de las principales causas de enfermedad respiratoria en caballos de todo el mundo. La prevención de la enfermedad es a través de la vacunación con vacunas a virus inactivado. La mayoría de las vacunas se producen en huevos embrionados, de los cuales los viriones son cosechados del líquido alantoideo e inactivados químicamente. Aunque este sistema ha servido bien durante años, el uso de huevos como sustrato para la producción de vacuna presenta varias desventajas bien reconocidas (costo, provisión de huevos, manejo de los residuos, rinde por huevo). El objetivo del presente trabajo fue evaluar preliminarmente un sistema de expresión en baculovirus como método de producción de hemoaglutinina recombinante (rHA) para ser utilizada como vacuna para la prevención de la influenza equina. Para ello el ectodominio de la hemaglutinina (la subunidad HA1) del virus de la influenza equina se expresó en células de insecto infectadas con un baculovirus recombinante. La expresión fue demostrada por SDS-PAGE e inmunoblotting. El método empleado fue capaz de producir gran cantidad de rHA1. En este estudio se obtuvieron 20 μg/ml (200 μg de HA1 purificada de 2,5x107 células infectadas). La respuesta inmune fue evaluada mediante la inmunización de ratones BALB/c. Los resultados preliminares demostraron que la proteína recombinante expresada en baculovirus genera una fuerte respuesta inmune en ratones, por lo tanto podría ser utilizada como antígeno para la producción de una vacuna a subunidades y en pruebas diagnósticas.(AU)


Assuntos
Animais , Feminino , Camundongos , Baculoviridae/metabolismo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/biossíntese , Vírus da Influenza A Subtipo H3N8/imunologia , Vacinas contra Influenza/biossíntese , Camundongos Endogâmicos BALB C , Vacinas Sintéticas/biossíntese
12.
Rev. argent. microbiol ; 45(4): 222-228, dic. 2013. ilus, tab
Artigo em Inglês | LILACS | ID: lil-708686

RESUMO

Equine influenza virus is a leading cause of respiratory disease in horses worldwide. Disease prevention is by vaccination with inactivated whole virus vaccines. Most current influenza vaccines are generated in embryonated hens' eggs. Virions are harvested from allantoic fluid and chemically inactivated. Although this system has served well over the years, the use of eggs as the substrate for vaccine production has several well-recognized disadvantages (cost, egg supply, waste disposal and yield in eggs). The aim of this study was to evaluate a baculovirus system as a potential method for producing recombinant equine influenza hemagglutinin to be used as a vaccine. The hemagglutinin ectodomain (HA1 subunit) was cloned and expressed using a baculovirus expression vector. The expression was determined by SDS-PAGE and immunoblotting. A high yield, 20 μg/ml of viral protein, was obtained from recombinant baculovirus-infected cells. The immune response in BALB/c mice was examined following rHA1 inoculation. Preliminary results show that recombinant hemagglutinin expressed from baculovirus elicits a strong antibody response in mice; therefore it could be used as an antigen for subunit vaccines and diagnostic tests.


El virus de la influenza equina es una de las principales causas de enfermedad respiratoria en caballos de todo el mundo. La prevención de la enfermedad es a través de la vacunación con vacunas a virus inactivado. La mayoría de las vacunas se producen en huevos embrionados, de los cuales los viriones son cosechados del líquido alantoideo e inactivados químicamente. Aunque este sistema ha servido bien durante años, el uso de huevos como sustrato para la producción de vacuna presenta varias desventajas bien reconocidas (costo, provisión de huevos, manejo de los residuos, rinde por huevo). El objetivo del presente trabajo fue evaluar preliminarmente un sistema de expresión en baculovirus como método de producción de hemoaglutinina recombinante (rHA) para ser utilizada como vacuna para la prevención de la influenza equina. Para ello el ectodominio de la hemaglutinina (la subunidad HA1) del virus de la influenza equina se expresó en células de insecto infectadas con un baculovirus recombinante. La expresión fue demostrada por SDS-PAGE e inmunoblotting. El método empleado fue capaz de producir gran cantidad de rHA1. En este estudio se obtuvieron 20 μg/ml (200 μg de HA1 purificada de 2,5x107 células infectadas). La respuesta inmune fue evaluada mediante la inmunización de ratones BALB/c. Los resultados preliminares demostraron que la proteína recombinante expresada en baculovirus genera una fuerte respuesta inmune en ratones, por lo tanto podría ser utilizada como antígeno para la producción de una vacuna a subunidades y en pruebas diagnósticas.


Assuntos
Animais , Feminino , Camundongos , Baculoviridae/metabolismo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/biossíntese , /imunologia , Vacinas contra Influenza/biossíntese , Camundongos Endogâmicos BALB C , Vacinas Sintéticas/biossíntese
13.
J Virol Methods ; 194(1-2): 102-6, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23948157

RESUMO

Honey bee mortality is a serious problem that beekeepers in Argentina have had to face during the last 3 years. It is known that the consequence of the complex interactions between environmental and beekeeping parameters added to the effect of different disease agents such as viruses, bacteria, fungi and parasitic mites may result in a sudden collapse of the colony. In addition, multiple viral infections are detected frequently concomitantly in bee colonies. The aim of this study was to establish a multiplex polymerase chain reaction method for rapid and simultaneous detection of the most prevalent bee viruses. This multiplex PCR assay will provide specific, rapid and reliable results and allow for the cost effective detection of a particular virus as well as multiple virus infections in a single reaction tube. This method could be a helpful tool in the surveillance of the most frequently found bee viruses and to study the dynamics and the interactions of the virus populations within colonies.


Assuntos
Abelhas/virologia , Reação em Cadeia da Polimerase Multiplex/métodos , Medicina Veterinária/métodos , Virologia/métodos , Vírus/isolamento & purificação , Animais , Argentina
14.
Rev Argent Microbiol ; 45(4): 222-8, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24401775

RESUMO

Equine influenza virus is a leading cause of respiratory disease in horses worldwide. Disease prevention is by vaccination with inactivated whole virus vaccines. Most current influenza vaccines are generated in embryonated hens' eggs. Virions are harvested from allantoic fluid and chemically inactivated. Although this system has served well over the years, the use of eggs as the substrate for vaccine production has several well-recognized disadvantages (cost, egg supply, waste disposal and yield in eggs). The aim of this study was to evaluate a baculovirus system as a potential method for producing recombinant equine influenza hemagglutinin to be used as a vaccine. The hemagglutinin ectodomain (HA1 subunit) was cloned and expressed using a baculovirus expression vector. The expression was determined by SDS-PAGE and immunoblotting. A high yield, 20µg/ml of viral protein, was obtained from recombinant baculovirus-infected cells. The immune response in BALB/c mice was examined following rHA1 inoculation. Preliminary results show that recombinant hemagglutinin expressed from baculovirus elicits a strong antibody response in mice; therefore it could be used as an antigen for subunit vaccines and diagnostic tests.


Assuntos
Baculoviridae/metabolismo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/biossíntese , Vírus da Influenza A Subtipo H3N8/imunologia , Vacinas contra Influenza/biossíntese , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Vacinas Sintéticas/biossíntese
15.
Rev. Argent. Microbiol. ; 45(4): 222-8, 2013 Oct-Dec.
Artigo em Espanhol | BINACIS | ID: bin-132757

RESUMO

Equine influenza virus is a leading cause of respiratory disease in horses worldwide. Disease prevention is by vaccination with inactivated whole virus vaccines. Most current influenza vaccines are generated in embryonated hens eggs. Virions are harvested from allantoic fluid and chemically inactivated. Although this system has served well over the years, the use of eggs as the substrate for vaccine production has several well-recognized disadvantages (cost, egg supply, waste disposal and yield in eggs). The aim of this study was to evaluate a baculovirus system as a potential method for producing recombinant equine influenza hemagglutinin to be used as a vaccine. The hemagglutinin ectodomain (HA1 subunit) was cloned and expressed using a baculovirus expression vector. The expression was determined by SDS-PAGE and immunoblotting. A high yield, 20Ag/ml of viral protein, was obtained from recombinant baculovirus-infected cells. The immune response in BALB/c mice was examined following rHA1 inoculation. Preliminary results show that recombinant hemagglutinin expressed from baculovirus elicits a strong antibody response in mice; therefore it could be used as an antigen for subunit vaccines and diagnostic tests.


Assuntos
Baculoviridae/metabolismo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/biossíntese , Vírus da Influenza A Subtipo H3N8/imunologia , Vacinas contra Influenza/biossíntese , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Vacinas Sintéticas/biossíntese
16.
Rev. argent. microbiol ; 43(4): 273-277, dic. 2011. ilus, tab
Artigo em Inglês | LILACS | ID: lil-634704

RESUMO

Equid herpesvirus 1 (EHV-1) infection has a signifcant economic impact on equine production, causing abortion, respiratory disease, neonatal death and neurological disorders. The identifcation of specifc EHV-1 genes related to virulence and pathogenicity has been the aim of several research groups. The purpose of the present study was to analyze different genomic regions of Argentinean EHV-1 strains and to determine their possible relationship with virulence or clinical signs. Twenty-fve EHV-1 Argentinean isolates recovered from different clinical cases between 1979 and 2007 and two reference strains were amplifed and sequenced. The sequence alignments were carried out using Clustal X version 1.92 and the putative amino acid sequences were deduced using Bio-Edit version 7.05. Minor changes were observed. No changes that could be involved in the different virulence in the mouse model of three EHV-1 Argentinean strains were found. No genetic variants were observed. The genomic regions analyzed are unsuitable for differentiation between abortigenic strains and those isolated from neonatal deaths.


La infección por Herpesvirus equino 1 (EHV-1) tiene un signifcativo impacto económico en la producción equina mundial al causar abortos, enfermedad respiratoria, muertes perinatales y desórdenes neurológicos. La identifcación de genes específcos relacionados con la virulencia y patogenicidad de este virus ha sido el propósito de varios grupos de investigación. En este trabajo se analizaron diferentes regiones genómicas de cepas argentinas de EHV-1 para determinar la posible relación entre la estructura genómica y la virulencia o los signos clínicos producidos. Veinticinco cepas aisladas de diferentes casos clínicos observados entre los años 1979 y 2007 y dos cepas de referencia fueron amplifcadas y secuenciadas. El alineamiento de las secuencias se realizó con el programa Clustal X versión 1.92; el programa Bio-Edit versión 7.05 permitió deducir la secuencia de aminoácidos. Solo se observaron cambios menores, no se encontraron variaciones que pudieran estar relacionadas con la diferencia de virulencia observada previamente en el modelo ratón. No se hallaron variantes genómicas. Las regiones genómicas analizadas no permitieron diferenciar cepas abortigénicas de aquellas aisladas de muertes neonatales.


Assuntos
Animais , Camundongos , Genoma Viral , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/genética , Doenças dos Cavalos/virologia , Sequência de Aminoácidos , Aborto Animal/epidemiologia , Aborto Animal/virologia , Argentina/epidemiologia , Sequência de Bases , DNA Viral/genética , Genes Virais , Cavalos , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , Herpesvirus Equídeo 1/classificação , Herpesvirus Equídeo 1/isolamento & purificação , Herpesvirus Equídeo 1/patogenicidade , Doenças dos Cavalos/epidemiologia , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Virulência/genética
17.
Biosci. j. (Online) ; 27(5): 826-829, sept./oct. 2011.
Artigo em Inglês | LILACS | ID: biblio-911874

RESUMO

The Przewalskii´s horse or Mongolian wild horse (Equus przewalskii, Poljakov, 1881) is presently the only species of wild horse in existence. Originally from Asia, it is, classified as in extremely high risk of extinction which puts the species in the seriously threatened category. The aim of this work was the preservation of tissues and the development of cell cultures from tissue samples obtained from a Przewalskii´s horse after its death. Biopsies of skin, skeletal and cardiac muscle, and ear cartilage were removed from a recently dead horse, added to Phosphate Buffer Saline (PBS) and refrigerated until processing. Some of the samples were frozen in liquid nitrogen and the other was grown as explants to generate fibroblast cell monolayers. The cell cultures obtained, were subsequently propagated with low passages, and frozen in liquid nitrogen, thus avoiding genetic and phenotypic alterations. The tissues and cell cultures were thawed to ascertain their viability by checking its progressive grow in a flask. It was not possible to obtain cultures from cardiac muscle. A bank of tissues and cells from the single Przewalskii´s horse that existed in Uruguay was generated, and can be used for scientific purposes and for the conservation of the species in the future


Assuntos
Preservação Biológica , Preservação de Tecido , Preservação de Amostras de Água , Espécies em Perigo de Extinção , Cavalos
18.
Rev Argent Microbiol ; 43(2): 84-6, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21731968

RESUMO

Honey bee mortality has recently been associated with Israeli acute paralysis virus (IAPV), a proposed etiological agent for a new syndrome known as Colony Collapse Disorder. Bees infected with this virus show shivering wings, progress into paralysis, and finally die outside the hive. During the last years, honey bee mortality became a serious problem for Argentinean beekeepers. We herein report the preliminary results of a survey carried out to detect IAPV in samples taken from several Argentine provinces, by using a reverse transcription Polymerase Chain Reaction assay. Our data indicate the existence of high frequency of IAPV in asymptomatic hives of Argentina.


Assuntos
Abelhas/virologia , Colapso da Colônia/virologia , Dicistroviridae/isolamento & purificação , Animais , Argentina/epidemiologia , Colapso da Colônia/epidemiologia , Feminino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estudos de Amostragem
19.
Rev. argent. microbiol ; 43(2): 84-86, jun. 2011. ilus, tab
Artigo em Inglês | LILACS | ID: lil-634676

RESUMO

Honey bee mortality has recently been associated with Israeli acute paralysis virus (IAPV), a proposed etiological agent for a new syndrome known as Colony Collapse Disorder. Bees infected with this virus show shivering wings, progress into paralysis, and finally die outside the hive. During the last years, honey bee mortality became a serious problem for Argentinean beekeepers. We herein report the preliminary results of a survey carried out to detect IAPV in samples taken from several Argentine provinces, by using a reverse transcription Polymerase Chain Reaction assay. Our data indicate the existence of high frequency of IAPV in asymptomatic hives of Argentina.


Recientemente la mortalidad de las abejas melíferas ha sido asociada al virus israelí de la parálisis aguda (IAPV), propuesto como agente etiológico del denominado síndrome de despoblamiento de las colmenas. Las abejas infectadas con este virus presentan temblores en las alas que progresan hasta convertirse en parálisis, y finalmente mueren fuera de la colmena. Durante los últimos años, la mortalidad de las abejas melíferas se ha transformado en un serio problema para los productores de miel de la Argentina. Nosotros informamos aquí los resultados preliminares de un estudio realizado para detectar IAPV en muestras de colmenas provenientes de varias provincias argentinas utilizando la técnica de transcripción reversa-reacción en cadena de la polimerasa. Nuestros datos indican la presencia de IAPV en un alto porcentaje de las colonias estudiadas.


Assuntos
Animais , Feminino , Abelhas/virologia , Colapso da Colônia/virologia , Dicistroviridae/isolamento & purificação , Argentina/epidemiologia , Colapso da Colônia/epidemiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estudos de Amostragem
20.
Rev Argent Microbiol ; 43(4): 273-7, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22274825

RESUMO

Equid herpesvirus 1 (EHV-1) infection has a significant economic impact on equine production, causing abortion, respiratory disease, neonatal death and neurological disorders. The identification of specific EHV-1 genes related to virulence and pathogenicity has been the aim of several research groups. The purpose of the present study was to analyze different genomic regions of Argentinean EHV-1 strains and to determine their possible relationship with virulence or clinical signs. Twenty-five EHV-1 Argentinean isolates recovered from different clinical cases between 1979 and 2007 and two reference strains were amplified and sequenced. The sequence alignments were carried out using Clustal X version 1.92 and the putative amino acid sequences were deduced using Bio-Edit version 7.05. Minor changes were observed. No changes that could be involved in the different virulence in the mouse model of three EHV-1 Argentinean strains were found. No genetic variants were observed. The genomic regions analyzed are unsuitable for differentiation between abortigenic strains and those isolated from neonatal deaths.


Assuntos
Genoma Viral , Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/genética , Doenças dos Cavalos/virologia , Aborto Animal/epidemiologia , Aborto Animal/virologia , Sequência de Aminoácidos , Animais , Argentina/epidemiologia , Sequência de Bases , DNA Viral/genética , Genes Virais , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , Herpesvirus Equídeo 1/classificação , Herpesvirus Equídeo 1/isolamento & purificação , Herpesvirus Equídeo 1/patogenicidade , Doenças dos Cavalos/epidemiologia , Cavalos , Camundongos , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Virulência/genética
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