ABSTRACT
Abstract This is a cross-sectional study to assess the presence of antibodies in ruminants against selected pathogens associated with reproductive disorders in cattle in four Brazilian states, including the zoonotic agent Coxiella burnetii. The used tests were Virus Neutralization Assay for IBR and BVD, Microscopic Agglutination Test for Leptospira spp., Indirect Fluorescent Antibody Test (IFAT) for C. burnetii and Toxoplasma gondii, and Enzyme-Linked Immunosorbent Assay for Neospora caninum and Trypanosoma vivax. Seropositivity for C. burnetii was 13.7% with titers from 128 to 131,072; 57.8% for BoHV-1, with titers between 2 and 1,024; 47.1% for BVDV-1a, with titers from 10 to 5,120; 89.2% for N. caninum; 50% for T. vivax; and 52.0% for Leptospira spp., with titers between 100 to 800 (the following serovars were found: Tarassovi, Grippotyphosa, Canicola, Copenhageni, Wolffi, Hardjo, Pomona and Icterohaemorrhagiae); 19.6% for T. gondii with titer of 40. This is the first study that has identified C. burnetii in cattle associated with BoHV and BVDV, N. caninum, Leptospira spp., T. gondii and T. vivax. Thus, future studies should be conducted to investigate how widespread this pathogen is in Brazilian cattle herds.
Resumo Este é um estudo transversal para avaliar a presença de anticorpos em ruminantes contra patógenos selecionados e associados a distúrbios reprodutivos em bovinos de quatro estados brasileiros, incluindo o agente zoonótico Coxiella burnetii. Os testes utilizados foram Teste de Vírus-Neutralização para BoHV e BVDV, teste de Aglutinação Microscópica para Leptospira spp., Reação de Imunofluorescência Indireta for C. burnetii e Toxoplasma gondii, e Ensaio de Imunoabsorção Enzimática para Neospora caninum e Trypanosoma vivax. A soropositividade para C. burnetii foi de 13,7% com títulos de 128 a 131.072; 57,8% para BoHV-1, com títulos entre 2 a 1.024; 47,1% para BVDV-1a, com títulos de 10 a 5.120; 89,2% para N. caninum; 50% para T. vivax; e 52,0% para Leptospira spp., com títulos entre 100 a 800 (sorovares encontrados: Tarassovi, Grippotyphosa, Canicola, Copenhageni, Wolffi, Hardjo, Pomona e Icterohaemorrhagiae) 19,6% para T. gondii com título de 40. Este é o primeiro estudo que evidencia a participação de C. burnetii em bovinos associada ao Vírus da Rinotraqueíte bovina infecciosa e da diarreia viral bovina, N. caninum, Leptospira spp., T. gondii e T. vivax em bovinos. Desta forma, futuros estudos devem ser conduzidos a fim de investigar o quão disseminado se encontra este patógeno em rebanhos bovinos brasileiros.
Subject(s)
Animals , Female , Cattle , Q Fever/veterinary , Trypanosomiasis, African/veterinary , Bovine Virus Diarrhea-Mucosal Disease/complications , Cattle Diseases/epidemiology , Toxoplasmosis, Animal/complications , Coccidiosis/veterinary , Leptospirosis/veterinary , Q Fever/complications , Q Fever/diagnosis , Q Fever/epidemiology , Toxoplasma/immunology , Trypanosomiasis, African/complications , Trypanosomiasis, African/diagnosis , Trypanosomiasis, African/epidemiology , Bovine Virus Diarrhea-Mucosal Disease/diagnosis , Bovine Virus Diarrhea-Mucosal Disease/epidemiology , Brazil/epidemiology , Agglutination Tests , Enzyme-Linked Immunosorbent Assay/veterinary , Cattle Diseases/microbiology , Cattle Diseases/parasitology , Cattle Diseases/virology , Seroepidemiologic Studies , Toxoplasmosis, Animal/diagnosis , Cross-Sectional Studies , Trypanosoma vivax , Coxiella burnetii/immunology , Coccidiosis/complications , Coccidiosis/diagnosis , Coccidiosis/epidemiology , Diarrhea Viruses, Bovine Viral/immunology , Neospora/immunology , Fluorescent Antibody Technique, Indirect/veterinary , Abortion, Veterinary , Endometritis/etiology , Infertility, Female/etiology , Leptospira/immunology , Leptospirosis/complications , Leptospirosis/diagnosis , Leptospirosis/epidemiologyABSTRACT
Abstract A large number of Brazilian zoos keep many endangered species of deer, however, very few disease surveillance studies have been conducted among captive cervids. Blood samples from 32 Brazilian deer (Blastocerus dichotomus, Mazama nana and Mazama americana) kept in captivity at Bela Vista Biological Sanctuary (Foz do Iguaçu, Brazil) were investigated for 10 ruminant pathogens, with the aims of monitoring deer health status and evaluating any potential zoonotic risk. Deer serum samples were tested for Brucella abortus, Leptospira (23 serovars), Toxoplasma gondii, Neospora caninum, bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, foot-and-mouth disease virus, western equine encephalitis virus, eastern equine encephalitis virus and Venezuelan equine encephalitis virus. Antibodies against T. gondii (15.6%), N. caninum (6.2%) and L. interrogans serogroup Serjoe (3.1%) were detected. The serological results for all other infectious agents were negative. The deer were considered to be clinically healthy and asymptomatic regarding any disease. Compared with studies on free-ranging deer, the prevalences of the same agents tested among the captive deer kept at the Sanctuary were lower, thus indicating good sanitary conditions and high-quality management practices at the zoo.
Resumo Um grande número de zoológicos brasileiros abriga espécies de cervídeos ameaçados de extinção, entretanto, estudos de vigilância de doenças em cervídeos de cativeiro são escassos. Amostras de sangue de 32 cervídeos brasileiros (Blastocerus dichotomus, Mazama nana e Mazama americana), mantidos em cativeiro no Refúgio Biológico Bela Vista (Foz do Iguaçu, Brasil), foram investigados para 10 patógenos de ruminantes, visando monitorar o estado de saúde dos cervídeos e avaliar a presença de agentes zoonóticos. As amostras de soro foram testadas para Brucella abortus, Leptospira (23 sorovares), Toxoplasma gondii, Neospora caninum, diarreia viral bovina, rinotraqueíte infecciosa bovina, febre aftosa, encefalomielite equina do oeste, encefalomielite equina do leste e encefalomielite equina venezuelana. Foram detectados anticorpos para T. gondii (15,6%), N. caninum (6,2%) e para L. interrogans sorogrupo Serjoe (3,1%). As sorologias apresentaram resultado negativo para as demais doenças. Os cervídeos foram considerados clinicamente sadios e assintomáticos para doenças. Comparados aos estudos de populações de vida livre, as soroprevalências para os mesmos agentes testados foram menores para os cervídeos mantidos no Refúgio, indicando as boas condições sanitárias e a qualidade das práticas de manejo no zoológico.
Subject(s)
Animals , Toxoplasma/immunology , Deer/immunology , Antibodies, Protozoan/blood , Neospora/immunology , Leptospira interrogans/immunology , Animals, Zoo/immunology , Brazil/epidemiology , Brucella abortus/immunology , Seroepidemiologic Studies , Toxoplasmosis, Animal/epidemiology , Coccidiosis/epidemiology , Diarrhea Viruses, Bovine Viral/immunology , Herpesvirus 1, Bovine/immunology , Foot-and-Mouth Disease Virus/immunology , Encephalitis Viruses/immunologyABSTRACT
Bovine viral diarrhea virus (BVDV) is an important cause of economic losses worldwide. E2 is an immunodominant protein and a promising candidate to develop subunit vaccines. To improve its immunogenicity, a truncated E2 (tE2) was fused to a single chain antibody named APCH, which targets to antigen-presenting cells. APCH-tE2 and tE2 proteins were expressed in the baculovirus system and their immunogenicity was firstly compared in guinea pigs. APCH-tE2 vaccine was the best one to evoke a humoral response, and for this reason, it was selected for a cattle vaccination experiment. All the bovines immunized with 1.5Ag of APCH-tE2 developed high levels of neutralizing antibodies against BVDV up to a year post-immunization, demonstrating its significant potential as a subunit vaccine. This novel vaccine is undergoing scale-up and was transferred to the private sector. Nowadays, it is being evaluated for registration as the first Argentinean subunit vaccine for cattle
El virus de la diarrea viral bovina (BVDV) es causante de importantes pérdidas económicas a nivel mundial. La proteína E2 es la inmunodominante del virus y es la candidata para desarrollar vacunas de subunidad. Para mejorar su inmunogenicidad, una versión truncada de la E2 (tE2) se fusionó a un anticuerpo de cadena simple (APCH), que se dirige a las células presentadoras de antígeno. Se expresaron las proteínas APCH-tE2 y tE2 en el sistema de baculovirus y su inmunogenicidad fue evaluada y comparada en cobayos; la proteína APCH-tE2 fue la que indujo la mejor respuesta humoral. Por dicha razón se la evaluó en bovinos utilizando 1,5µg de antígeno. Los animales presentaron altos títulos de anticuerpos neutralizantes contra BVDV hasta un año posinmunización. Esta nueva vacuna está en proceso de escalado y se transfirió al sector privado. Actualmente se está evaluando para su registro como la primera vacuna argentina de subunidad para bovinos
Subject(s)
Animals , Cattle , Guinea Pigs , Diarrhea Viruses, Bovine Viral/immunology , Vaccines, Subunit/biosynthesis , Antigen-Presenting Cells/drug effects , Baculoviridae/immunology , Immunization/veterinary , Adenovirus E2 Proteins/immunology , Diarrhea Viruses, Bovine Viral/drug effects , Antibodies, Neutralizing/analysisABSTRACT
A serological survey was carried out to determine the prevalence rate of bovine viral diarrhea virus [BVDV] infections in water buffalo [Bubalusbubalis] in Ahvaz, which is the center of the Khouzestan province in Iran. For this purpose, blood samples were taken from 310 slaughtered buffaloes at the abattoir. Sera were tested via the serum neutralization test. Serum neutralization was performed by National Animal Diseases Laboratory [NADL], in order to isolate the genotype 1 strain of bovine viral diarrhea virus. The results indicate that 105 [33.9%] buffaloes had antibodies to BVDV. The prevalence of infection in females and males were 39.5% and 22.78%, respectively, and statistical analysis showed that this difference was significant. Although there was a non-significant difference between heifers and males, the difference between cows and bulls was highly significant
Subject(s)
Animals , Male , Female , Buffaloes , Diarrhea Viruses, Bovine Viral/immunologyABSTRACT
The bovine viral diarrhea virus (BVDV) infection control should be based on elimination of persistently infected animals and on immunization through vaccination, to prevent fetal infection. However, the efficacy of inactivated BVDV vaccines is variable due to its low immunogenicity. This study evaluated the humoral immune response against homologous and heterologous strains of 7 inactivated BVDV vaccines, in bovines and two experimental models (ovine and guinea pig) which might be used to test candidate vaccines. Vaccines formulated with BVDV Singer, Oregon, NADL, and multivalent, induced seroconversion in the three animal models studied, reaching antibody titres higher than 2. Vaccine containing 125C -genotype 2- only induced a low antibody response in ovine, while VS-115 NCP vaccine was not immunogenic. Furthermore, bovine sera at 60 dpv presented homologous as well as heterologous antibody response, indicating a high degree of cross-reactivity among the strains studied. However, when bovine sera were tested against the Argentine field strain 00-693, they showed the lowest levels of cross-reactivity, suggesting the need of continued surveillance to identify and characterize emerging field BVDV strains. Finally, optimal correlations between bovine-ovine and bovine-guinea pig models were observed, indicating that two alternative species could replace bovines when testing the immunogenicity of BVDV candidate vaccines.
El control del virus de la diarrea viral bovina (VDVB) se basa en la eliminación de animales persistentemente infectados, y la inmunización de hembras para prevenir infecciones fetales. La eficiencia de estas vacunas es variable por su baja inmunogenicidad. Se evaluó la respuesta inmune humoral contra virus homólogos y heterólogos de 7 vacunas experimentales inactivadas del VDVB en bovinos y en dos modelos experimentales (ovinos y cobayos). Las vacunas conteniendo VDVB Singer, Oregon, NADL y polivalentes indujeron seroconversión en los tres modelos y se alcanzaron títulos de anticuerpos mayores de 2. La vacuna con VDVB genotipo 2 VS-115, NCP, no resultó inmunogénica. La vacuna genotipo 2 125C sólo indujo baja respuesta humoral en ovinos, mientras que la VS-115, NCP, no resultó inmunogénica. En bovinos se determinó la respuesta a virus homólogos y heterólogos a 60 dpv, lo que indica un alto grado de inmunidad cruzada entre la mayoría de las cepas estudiadas. Cuando los sueros bovinos fueron ensayados con la cepa de campo de Argentina 00-693, los niveles de reacción cruzada fueron más bajos; esto sugiere la necesidad de una vigilancia epidemiológica sostenida a fin de identificar y caracterizar las cepas emergentes del VDVB. La óptima correlación en el modelo bovino-ovino y bovino-cobayo indica su utilidad para evaluar la inmunogenicidad de vacunas inactivadas de VDVB.
Subject(s)
Animals , Cattle , Guinea Pigs , Diarrhea Viruses, Bovine Viral/immunology , Viral Vaccines/immunology , Antibodies, Viral/biosynthesis , Antibodies, Viral/immunology , Dose-Response Relationship, Immunologic , Diarrhea Viruses, Bovine Viral/genetics , Genotype , Models, Animal , Neutralization Tests , Sheep , Species Specificity , Vaccines, Inactivated/immunologyABSTRACT
Two recombinant baculoviruses were produced in order to obtain a bovine viral diarrhea virus (BVDV) immunogen: AcNPV/E2 expressing E2 glycoprotein, and AcNPV/E0E1E2 expressing the polyprotein region coding for the three structural proteins of BVDV (E0, E1, and E2). Mice were immunized with Sf9 cells infected with the recombinant baculoviruses in a water in oil formulation and the production of neutralizing antibodies was evaluated. Since E2 elicited higher neutralizing antibody titers than E0-E1-E2 polyprotein, it was selected to immunize cattle. Calves received two doses of recombinant E2 vaccine and were challenged with homologous BVDV 37 days later. The recombinant immunogen induced neutralizing titers which showed a mean value of 1.5 ± 0.27 on the day of challenge and reached a top value of 3.36 ± 0.36, 47 days later (84 days post-vaccination). On the other hand, sera from animals which received mock-infected Sf9 cells did not show neutralizing activity until 25 days post-challenge (62 days post-vaccination), suggesting that these antibodies were produced as a consequence of BVDV challenge. Even when no total protection was observed in cattle, in vitro viral neutralization assays revealed that the recombinant immunogen was able to induce neutralizing antibody synthesis against the homologous strain as well as against heterologous strains in a very efficient way.
Subject(s)
Animals , Cattle , Mice , Bovine Virus Diarrhea-Mucosal Disease/prevention & control , Diarrhea Viruses, Bovine Viral/immunology , Viral Envelope Proteins/immunology , Viral Vaccines/immunology , Antibodies, Viral/blood , Antibodies, Viral/immunology , Bovine Virus Diarrhea-Mucosal Disease/immunology , Neutralization Tests , Recombinant Proteins/immunology , Time Factors , Vaccines, Synthetic/immunologyABSTRACT
A resposta sorológica induzida por três vacinas comerciais inativadas contra o vírus da Diarréia Viral Bovina (BVDV) foi avaliada em bovinos imunizados três vezes (dias 0, 30 e 180) e testados a diferentes intervalos após a vacinaçäo. Trinta dias após a segunda vacinaçäo, 74,5 por cento (70/94) dos animais apresentavam anticorpos neutralizantes contra o BVDV-1 e 52,1 por cento (49/94) contra o BVDV-2. Os títulos médios (GMT) e o número de animais reagentes contra o BVDV-1 eram de 109,3 (32/36); 54,6 (22/28) e 25,5 (16/30) para as vacinas A, B e C, respectivamente; e de 19 (27/36), 42,3 (12/28) e 18,4 (10/30) contra o BVDV-2. Os títulos reduziram-se aos 180 dias, sendo que 31,9 por cento (30/94) dos animais já näo apresentavam atividade neutralizante frente ao BVDV-1 e 63,8 por cento (60/94) frente ao BVDV-2. Nesta data, os títulos médios e o número de animais positivos frente ao BVDV-1 eram de 28,3 (30/36), 28,3 (20/28) e 16,1 (14/30) e frente ao BVDV-2 de 16,8 (18/36), 21,6 (10/28) e 28,3 (6/30) para as vacinas A, B e C, respectivamente. Após o reforço (dia 180), os títulos médios contra o BVDV-1 aumentaram significativamente nos três grupos vacinais e contra o BVDV-2 apenas no grupo A. Trinta dias após, os títulos médios e o número de reagentes contra o BVDV-1 eram de 104,8 (23/24), 50,3 (24/26) e 43,7 (24/28) e contra o BVDV-2 de 33,4 (23/24), 23,3 (22/26) e 15,7 (22/28) para as vacinas A, B e C. Os títulos contra o BVDV-1 no dia 210 foram estatisticamente superiores aos títulos contra o BVDV-2 nos três grupos vacinais. O soro de alguns animais positivos de cada grupo foi testado frente a quatro amostras brasileiras de BVDV-1 e duas de BVDV-2. Além dos títulos baixos a moderados, os testes de neutralizaçäo cruzada revelaram variaçöes marcantes na atividade neutralizante frente a isolados de campo antigenicamente diferentes. Esses resultados demonstram que a vacinaçäo näo induziu uma resposta sorológica de magnitude e duraçäo adequadas na maioria dos animais, principalmente frente à grande diversidade antigênica das amostras de BVDV.
Subject(s)
Animals , Bovine Virus Diarrhea-Mucosal Disease/immunology , Diarrhea Viruses, Bovine Viral/immunology , Cattle , Serologic Tests/veterinaryABSTRACT
A resposta sorológica e proteçäo fetal conferida por três vacinas inativadas contra o vírus da diarréia viral bovina (BVDV) (vacinas A, B e C) foram avaliadas através de vacinaçäo e posterior desafio de ovelhas prenhes com amostras brasileiras de BVDV-1 e BVDV-2. Níveis baixos a moderados de anticorpos neutralizantes anti-BVDV-1 foram detectados na maioria dos animais (45/47) aos 30 dias após a segunda dose vacinal (títulos médios geométricos [GMT] de 124,7; 74,6 e 26,7 para as vacinas A, B e C, respectivamente). Em contraste, atividade neutralizante anti-BVDV-2 näo foi detectada em vários animais (12/47) e foi de magnitude inferior nos três grupos vacinais (GMTs 19,1; 14,1 e 15,1). Os títulos médios de anticorpos reduziram-se significativamente no dia 180, sendo que vários animais já näo apresentavam atividade neutralizante detectável frente ao BVDV-1 (grupo B=1/19; C=8/14) e principalmente frente ao BVDV-2 (A=7/14; B=13/19; C=13/14). Nessa data, os títulos médios de anticorpos contra as amostras utilizadas no desafio eram de 91,9; 15,1 e 60,6 (SV-126.8, BVDV-1) e de 10; <10 e 28,3 (SV-260, BVDV-2) nos grupos A, B e C, respectivamente. Nos três grupos vacinais, os níveis de anticorpos neutralizantes contra essas amostras näo foram sufucientes para prevenir a replicaçäo, disseminaçäo virêmica e transmissäo transplacentária dos vírus aos fetos. O vírus foi detectado no sangue e nos fetos de todas as ovelhas vacinadas com as vacinas A (10/10), B (9/9) e C (8/8). Esses resultados demonstraram que as vacinas testadas induziram níveis moderados a baixos de anticorpos neutralizantes contra o BVDV-1 e, principalmente contra o BVDV-2 na maioria dos animais; e que esses níveis näo foram suficientes para prevenir a infecçäo fetal frente ao desafio com amostras de BVDV-1 e BVDV-2. Adicionalmente, os resultados confirmam a adequaçäo de ovelhas prenhes para estudos de proteçäo fetal por vacinas contra o BVDV.
Subject(s)
Animals , Female , Sheep Diseases/immunology , Viral Vaccines , Diarrhea Viruses, Bovine Viral/immunology , SheepABSTRACT
The BVDV glycoproteins gp48 and gp53 were expressed in the baculovirus eukaryotic system. Both recombinant proteins were recognized in western blot analysis by monoclonal antibodies and polyclonal serum. Immunofluorescent test demonstrated that gp53 was localized on the cell surface whereas gp48 was in the cytoplasm. The expressed proteins were extracted by non-denaturing detergent treatment. Rabbit antiserum raised against gp53 recombinant protein efficiently neutralized the virus. These results demonstrate that the recombinant proteins have immunological properties similar to those of the native viral protein and that they can be useful as diagnostic reagents.
Subject(s)
Animals , Cattle , Male , Rabbits , Viral Envelope Proteins/isolation & purification , Diarrhea Viruses, Bovine Viral/chemistry , Blotting, Western , Cell Line , Immune Sera , Kidney , Nucleopolyhedroviruses , Viral Envelope Proteins/genetics , Viral Envelope Proteins/immunology , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/isolation & purification , Spodoptera , Testis/cytology , Transfection , Genetic Vectors/genetics , Diarrhea Viruses, Bovine Viral/genetics , Diarrhea Viruses, Bovine Viral/immunologyABSTRACT
The tissue distribution and cellular localization of viral antigens in three cattle with persistent bovine viral diarrhea virus (BVDV) infection was studied. In three cases, necropsy findings of oral ulcers, abmasal ulcers and necrosis of Peyer's patches were suspected have been caused by BVDV infection. Non-cytopathic BVDV was isolated from a tissue pool of liver, kidneys and spleen. Immunohistochemical detection of BVDV showed that BVDV antigens were detected in both epithelial and nonepithelial cells in all examined organs, including the gastrointestinal tract, liver, pancreas, lung, lymphatic organs (spleen, lymph nodes), adrenal gland, ovary, uterus, and the mammary gland. These findings support the hypothesis that animals with persistent BVDV infection spread BVDV through all routes, and that infertility in BVDV infection is associated with the infection of BVDV in the ovaries and uteri.
Subject(s)
Animals , Cattle , Female , Adrenal Glands/pathology , Antigens, Viral/isolation & purification , Bovine Virus Diarrhea-Mucosal Disease/pathology , Diarrhea Viruses, Bovine Viral/immunology , Digestive System/pathology , Immunohistochemistry/veterinary , Infertility, Female/virology , Kidney/pathology , Lung/pathology , Lymphatic System/pathology , Mammary Glands, Animal/virology , Ovary/pathology , Uterus/pathologyABSTRACT
Three Brazilian isolates of bovine viral diarrhea virus (BVDV), antigenically distinct from the standard North American isolates, were selected to immunize BALB/c mice in order to obtain hybridoma cells secreting anti-BVDV monoclonal antibodies (mAbs). Two hybridoma clones secreting mAbs, reacting specifically with BVDV-infected cells (mAbs 3.1C4 and 6.F11), were selected after five fusions and screening of 1001 hypoxanthine-aminopterin-thymidine-resistant clones. These mAbs reacted in an indirect fluorescent antibody (IFA) assay with all 39 South and North American BVDV field isolates and reference strains available in our laboratory, yet failed to recognize other pestiviruses, namely the hog cholera virus. The mAbs reacted at dilutions up to 1:25,600 (ascitic fluid) and 1:100 (hybridoma culture supernatant) in IFA and immunoperoxidase (IPX) staining of BVDV-infected cells but only mAb 3.1C4 neutralized virus infectivity. Furthermore, both mAbs failed to recognize BVDV proteins by IPX in formalin-fixed paraffin-embedded tissues and following SDS-PAGE and immunoblot analysis of virus-infected cells, suggesting they are probably directed to conformational-type epitopes. The protein specificity of these mAbs was then determined by IFA staining of CV-1 cells transiently expressing each of the BVDV proteins: mAb 3.1C4 reacted with the structural protein E2/gp53 and mAb 6.F11 reacted with the structural protein E1/gp25. Both mAbs were shown to be of the IgG2a isotype. To our knowledge, these are the first mAbs produced against South American BVDV isolates and will certainly be useful for research and diagnostic purposes
Subject(s)
Animals , Mice , Cattle , Antibodies, Monoclonal/biosynthesis , Antibodies, Monoclonal/immunology , Diarrhea Viruses, Bovine Viral/immunology , Hybridomas , Antigenic Variation , Diarrhea Viruses, Bovine Viral/genetics , Diarrhea Viruses, Bovine Viral/isolation & purification , Fluorescent Antibody Technique, Indirect , Genotype , Horses , Mice, Inbred BALB C , Recombinant ProteinsABSTRACT
Nineteen Brazilian isolates of bovine viral diarrhea virus (BVDV) were characterized antigenically with a panel of 19 monoclonal antibodies (mAbs) (Corapi WV, Donis RO and Dubovi EJ (1990) American Journal of Veterinary Research, 55: 1388-1394). Eight isolates were further characterized by cross-neutralization using sheep monospecific antisera. Analysis of mAb binding to viral antigens by indirect immunofluorescence revealed distinct patterns of reactivity among the native viruses. Local isolates differed from the prototype Singer strain in recognition by up to 14 mAbs. Only two mAbs - one to the non-structural protein NS23/p125 and another to the envelope glycoprotein E0/gp48 - recognized 100 per cent of the isolates. No isolate was recognized by more than 14 mAbs and twelve viruses reacted with 10 or less mAbs. mAbs to the major envelope glycoprotein E2/gp53 revealed a particularly high degree of antigenic variability in this glycoprotein. Nine isolates (47.3 per cent) reacted with three or less of 10 E2/gp53 mAbs, and one isolate was not recognized by any of these mAbs. Virus-specific antisera to eight isolates plus three standard BVDV strains raised in lambs had virus-neutralizing titers ranging from 400 to 3200 against the homologous virus. Nonetheless, many antisera showed significantly reduced neutralizing activity when tested against heterologous viruses. Up to 128-fold differences in cross-neutralization titers were observed for some pairs of viruses. When the coefficient of antigenic similarity (R) was calculated, 49 of 66 comparisons (74.24 per cent) between viruses resulted in R values that antigenically distinguish strains. Moreover, one isolate had R values suggesting that it belongs to a distinct serologic group. The marked antigenic diversity observed among Brazilian BVDV isolates should be considered when planning diagnostic and immunization strategies.
Subject(s)
Antibodies, Monoclonal , Antigenic Variation , Diarrhea Viruses, Bovine Viral/immunology , Brazil , Diarrhea Viruses, Bovine Viral/isolation & purification , Neutralization TestsABSTRACT
76 bovine serum samples were collected from different farms and tested for the presence of antibodies to BVD virus. For this purpose, the SPA-agllutination test has been introduced as a new approach in comparison with the serum neutralization test [SNT] and the immunodiffusion [ID] technique. Therefore, the samples have been categorized in four groups according to their titers in the SNI: 1st group included positive sera with high antibody titer [32 to 64]; 2nd group included positive sera with medium antibody titer [8 to 16]; 3rd group contained cytotoxic samples and the 4th group comprised all negative samples. The obtained data could prove that the SPA test as a binding assay is relatively more sensitive than the SNT in detecting antibodies to BBVD virus giving end titers ranged from 16 to 128. The SPA test could also detect anti-BVD antibodies in cytotoxic serum samples. Although the ID test is a simple technique for detection of antibodies yet it was of relative lower sensitivity than the SNT and SPA agglutination tests, and showed itself in this work non-suitable for screening of antibodies to BVD virus