<p>The human coronavirus NL63 was discovered in 2004 by a team of researchers in Amsterdam. Since its discovery it has been shown to have worldwide spread and affects mainly children, aged 0-5 years old, the immunocompromised and the elderly. Infection with HCoV-NL63 commonly results in mild upper respiratory tract infections and presents as the common cold, with symptoms including fever, cough, sore throat and rhinorrhoea. Lower respiratory tract findings are less common but may develop into more serious complications including bronchiolitis, pneumonia and croup. The primary function of the HCoV-NL63 nucleocapsid (N) protein is the formation of theprotective ribonucleocapsid core. For this particle to assemble, the N-protein undergoes N-N dimerization and then interacts with viral RNA. Besides the primary structural role of the Nprotein, it is also understood to be involved in viral RNA transcription, translation and replication, including several other physiological functions. The N-protein is also highly antigenic and elicits a strong immune response in infected patients. For this reason the N-protein may serve as a target for the development of diagnostic assays. We have used bioinformatic analysis to analyze the HCoV-NL63 N-protein and compared it to coronavirus N-homologues. This bioinformatic analysis provided the data to generate recombinant clones for expression in a bacterial system. We constructed recombinant clones of the N-protein of SARS-CoV and HCoV-NL63 and synthesized truncated clones corresponding to the N- and C-terminal of the HCoV-NL63 N-protein. These heterologously expressed proteins will serve the basis for several post-expression studies including characterizing the immunogenic epitope of the N-protein as well identifying any antibody crossreactivity between coronavirus species.</p>

<p>The human coronavirus NL63 was discovered in 2004 by a team of researchers in Amsterdam. Since its discovery it has been shown to have worldwide spread and affects mainly children, aged 0-5 years old, the immunocompromised and the elderly. Infection with HCoV-NL63 commonly results in mild upper respiratory tract infections and presents as the common cold, with symptoms including fever, cough, sore throat and rhinorrhoea. Lower respiratory tract findings are less common but may develop into more serious complications including bronchiolitis, pneumonia and croup. The primary function of the HCoV-NL63 nucleocapsid (N) protein is the formation of theprotective ribonucleocapsid core. For this particle to assemble, the N-protein undergoes N-N dimerization and then interacts with viral RNA. Besides the primary structural role of the Nprotein, it is also understood to be involved in viral RNA transcription, translation and replication, including several other physiological functions. The N-protein is also highly antigenic and elicits a strong immune response in infected patients. For this reason the N-protein may serve as a target for the development of diagnostic assays. We have used bioinformatic analysis to analyze the HCoV-NL63 N-protein and compared it to coronavirus N-homologues. This bioinformatic analysis provided the data to generate recombinant clones for expression in a bacterial system. We constructed recombinant clones of the N-protein of SARS-CoV and HCoV-NL63 and synthesized truncated clones corresponding to the N- and C-terminal of the HCoV-NL63 N-protein. These heterologously expressed proteins will serve the basis for several post-expression studies including characterizing the immunogenic epitope of the N-protein as well identifying any antibody crossreactivity between coronavirus species.</p>

Orientador: Hélio José Montassier / Banca: Camillo Del Cistia Andrade / Banca: Janete Apparecida Desidério Sena. / Resumo: Anticorpos monoclonais se constituem na base de vários testes usados na detecção e na identificação de antígenos. Nesse contexto, tais imuno-reagentes têm sido extensivamente empregados na identificação de estirpes virais envolvidas na etiologia de surtos de bronquite infecciosa a campo, permitindo o aperfeiçoamento das técnicas de detecção e caracterização antigênica do vírus da bronquite infecciosa das galinhas (VBI). No presente estudo, uma biblioteca de fragmentos de anticorpos de galinha originalmente preparada por "phage display" contra a estirpe vacinal (H120) do VBI, foi usada para a seleção de fragmentos de anticorpos recombinantes com reatividade cruzada para as estirpes heterólogas IBVPR01, IBVPR05, isoladas de surtos a campo no Brasil e SE-17, isolada nos Estados Unidos. Após três ciclos de "panning", foi identificado pelo ELISA um conjunto de 15 anticorpos scFv expressos em fagos e com reatividade cruzada para essas mesmas estirpes do VBI. A análise por Western-blotting revelou que três desses clones apresentavam fagos expressando fragmentos de anticorpos monoclonais com reatividade cruzada para a nucleoproteína N das três estirpes do VBI e também para a forma recombinante dessa nucleoproteína derivada da estirpe M41. Concluindo, os fragmentos de anticorpos monoclonais recombinantes scFv-N produzido em fagos interagem com um epítopo mais conservado da proteína N do VBI e apresentam um grande potencial para utilização na detecção e no diagnóstico direto desse vírus e no estudo de evolução de variantes desse vírus. / Abstract: Monoclonal antibodies are the basis of various techniques used for antigen detection or characterization, and their use is specially recommended for the identification of viral strains involved in the etiology of outbreaks of infectious bronchitis, because these antibodies are homogeneous, highly specific and fully characterized, allowing the improvement of detection of immunological techniques and antigenic characterization of avian infectious bronchitis virus strains (IBV). We used a phage display library prepared previously against the IBV vaccine strain (H120) for the selection of new scFv antibody fragments reacting with heterologous IBV strains isolated from outbreaks in Brazil (IBVPR01, IBVPR05) and USA (SE-17). After three cycles of panning a set of 15 scFv antibodies was expressed in phages and exhibited crossreaction in ELISA with these three viral strains. Western-blotting analysis showed that three of this clone set were expressing scFv specific for the nucleoprotein of these IBV strains, as well as to the recombinant form of this protein derived from M41 strain of IBV. In conclusion, the recombinant fragments of monoclonal antibodies expressed by phage-display technique have a great potential for future use in immunodiagnostic techniques and study the evolution of variant strains of this virus. / Mestre

Infectious bovine keratoconjunctivitis (IBK) is the main ocular disease of cattle. Highly contagious, it is responsible for significant economic losses of livestock worldwide. Moraxella bovis is recognized as the primary agent of the disease in cattle. However, the occurrence of other similar species, including M. ovis and M. bovoculi, recently described, have been common in outbreaks of the disease and are suspected to be causally related to the limited success of preventive measures, especially the use of bacterins containing only strains of M. bovis. This dissertation describes the antigenic characterization of M. bovis, M. bovoculi and M. ovis strains and a commercial vaccine by cross-reactivity using flow cytometry analysis and identification of immunodominant conserved antigens by Western blotting. Antisera against strains of the three species were obtained from immunization of New Zealand rabbits and challenged before a panel of strains isolated from cattle and sheep clinically affected by the disease between 1983 and 2013 as well as reference strains of each species. Field strains of M. bovoculi (Mbv2 and Mbv3) recognized satisfactorily all heterologous strains. However, Mbv3 (M. bovoculi), Mov2 (M. ovis) and Mov3 (M. ovis) strains stood out as the most intense recognition strains of their respective species and suggest that species-specific antigens play an important role in the host immune response. The association of reactivity percentage with the antigenic profiles, evidenced by Western blotting analysis, indicates that the immune response induced by Moraxella spp. appears to be mediated by multiple surface antigens of which many are shared between the three species. Among 32 different proteins identified, 22 (68.7%) were recognized by at least one antiserum in the protein extracts of all strains analyzed by Western blotting. Our study suggests (1) the selection and combination of M. bovis, M. bovoculi and M. ovis strains to be used in the composition of antigenic unit vaccines as IBK control strategy and (2) the use of flow cytometry as the most appropriate methodology for this selection. / Ceratoconjuntivite infecciosa bovina (CIB) é a principal doença ocular de bovinos. Altamente contagiosa, é responsável por significativas perdas econômicas para a pecuária no mundo inteiro. Moraxella bovis é, reconhecidamente, o agente primário da enfermidade em bovinos. Contudo, a ocorrência de outras espécies do gênero, incluindo M. ovis e M. bovoculi, recentemente descrita, têm sido comuns em surtos da doença e são suspeitas de serem causalmente associadas ao sucesso limitado de medidas preventivas, sobretudo do uso de bacterinas contendo apenas cepas de M. bovis. Esta dissertação descreve a caracterização antigênica de cepas de M. bovis, M. bovoculi e M. ovis e de uma vacina comercial através da análise de reatividade cruzada por citometria de fluxo e da identificação de antígenos imunodominantes e conservados através de Western blotting. Antissoros contra cepas das três espécies foram obtidos a partir da imunização de coelhos Nova Zelândia e desafiados frente a um painel de cepas isoladas de bovinos e ovinos clinicamente acometidos pela doença entre 1983 e 2013 assim como cepas de referência de cada espécie. Cepas de campo de M. bovoculi (Mbv2 e Mbv3) reconheceram satisfatoriamente todas as cepas heterólogas. Contudo, as cepas Mbv3 (M. bovoculi), Mov2 (M. ovis) e Mov3 (M. ovis) destacaram-se pela maior intensidade de reconhecimento de cepas de suas respectivas espécies e sugerem que antígenos espécie-específicos desempenham importante papel na resposta imune do hospedeiro. A associação dos percentuais de reatividade aos perfis antigênicos evidenciados através da análise por western blotting indica que a resposta imune induzida por Moraxella spp. parece ser mediada por múltiplos antígenos de superfície dos quais, diversos são compartilhados entre as três espécies. Entre 32 diferentes proteínas detectadas, 22 (68,7%) foram reconhecidas por pelo menos um antissoro nos extratos protéicos de todas as cepas analisadas por western blotting. Nosso estudo sugere a seleção e combinação de cepas de M. bovis, M. bovoculi e M. ovis circulantes para composição da unidade antigênica de vacinas como estratégia de controle de IBK e a utilização de citometria de fluxo como metodologia mais adequada para esta seleção.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / The human coronavirus NL63 was discovered in 2004 by a team of researchers in Amsterdam. Since its discovery it has been shown to have worldwide spread and affects mainly children, aged 0-5 years old, the immunocompromised and the elderly. Infection with HCoV-NL63 commonly results in mild upper respiratory tract infections and presents as the common cold, with symptoms including fever, cough, sore throat and rhinorrhoea. Lower respiratory tract findings are less common but may develop into more serious complications including bronchiolitis, pneumonia and croup. The primary function of the HCoV-NL63 nucleocapsid (N) protein is the formation of theprotective ribonucleocapsid core. For this particle to assemble, the N-protein undergoes N-N dimerization and then interacts with viral RNA. Besides the primary structural role of the Nprotein, it is also understood to be involved in viral RNA transcription, translation and replication, including several other physiological functions. The N-protein is also highly antigenic and elicits a strong immune response in infected patients. For this reason the N-protein may serve as a target for the development of diagnostic assays. We have used bioinformatic analysis to analyze the HCoV-NL63 N-protein and compared it to coronavirus N-homologues. This bioinformatic analysis provided the data to generate recombinant clones for expression in a bacterial system. We constructed recombinant clones of the N-protein of SARS-CoV and HCoV-NL63 and synthesized truncated clones corresponding to the N- and C-terminal of the HCoV-NL63 N-protein. These heterologously expressed proteins will serve the basis for several post-expression studies including characterizing the immunogenic epitope of the N-protein as well identifying any antibody crossreactivity between coronavirus species. / South Africa

While a decrease of wild felid population has led to disruption of conservation programme, recent studies have shown the importance of immune regulation for determining health outcomes and co-infection. Immunoglobulin G is important for detecting and evaluating responses to infectious diseases and vaccination. But, there is limited information on felid immunoglobulins and their role for functional immunity. This study aimed at isolating and characterizing lion’s immunoglobulin G. Lions’ sera (n = 68) were processed using the MagReSyn® magnetic beads and the final protein concentration was determined using the Xpose™ Trinean Spectrophotometer. The cross-reactivity of goat anti-cat immunoglobulin with sera of lions and other species was analysed using ELISA. High cross-reactivity was observed in lions ranging from 87.7 to 100%, and low reactivity with rhino (22.4%) followed by chicken (0.01%). The protein concentration from purified sera yielded 39.09 mg/ml. Molecular weight of lion IgG 150-160 kDa was detected with both chains at 54-56 kDa and 24-26 kDa on SDS PAGE. These results indicate a potential aid in developing serological tools to monitor exposure to micro-organisms of lions. / Agriculture and  Animal Health / M. Sc. (Agriculture)

Os tripanosomatídeos inferiores, que infectam plantas e insetos, apresentam propriedades bioquímicas e moleculares similares a Leishmania sp. e Trypanosoma cruzi. Similaridades antigênicas entre estes parasitas são conhecidas à muito tempo, mas somente alguns poucos estudos comparativos sobre a imunorreatividade humoral cruzada foram descritos. No presente trabalho nós analisamos a imunorreatividade cruzada de extratos antigênicos totais de oito tripanosomatídeos inferiores por ELISA, com soros de humanos e cães infectados com T. cruzi e Leishmania sp. Segundo as positividades e dados de reatividade média para ELISA os tripanosomatídeos inferiores foram divididos em dois grupos. ELISA-G1 compreendeu 4 parasitas para os quais se obtiveram 100% de positividade e elevadas médias de absorbância, similar aos dados obtidos para L. chagasi para hospedeiros humanos e cães. Nos casos humanos, soros de pacientes chagásicos crônicos apresentaram 100% de positividade somente para o ELISA com T. cruzi, sem diferenças entre os tripanosomatídeos inferiores. Por outro lado amostras de doenças não relacionadas apresentaram baixa reatividade cruzada com os tripanosomatídeos inferiores. Apesar da sua posição taxonômica em várias sessões e sua antiga divergência estes resultados mostraram semelhança antigênica entre os tripanosomatídeos inferiores e os patogênicos, e podem ser uma fonte alternativa de antígenos para a detecção de anticorpos principalmente em casos de leishmaniose visceral. / \"Lower trypanosomatids\", that infect plant and insect, present biochemical and molecular similarities to Leishmania sp. and Trypanosoma cruzi. Antigenic similarities between those parasites are known for a long time, but only few comparative investigations about immune humoral cross-reaction were described. In the current work we analyze the cross-immunoreactivity of crude extract from eight lower trypanosomatids by ELISA, with human and dog host samples infected with T. cruzi and Leishmania sp. ELISA positivity and data of mean title of human or dog visceral leishmaniasis cases, the lower trypanosomatids were divided in two groups. ELISA-G1 comprised by 4 parasites resulted in 100% positivity and high mean of absorbance, similar data to those obtained with L. chagasi, with human or dog hosts. In human cases, Chagas disease chronic cases showed 100% positive only with ELISA performed with T. cruzi, with no differences among lower trypanosomatids. Otherwise samples with other non correlated disease presented low cross-reaction with lower trypanososmatids. In spite of, their taxonomic position in various sections and their old divergence these results showed a strong antigenic similarity between pathogenic and lower trypanosomatids, and could be an alternative source of antigen for the detection of antibodies against host mainly with visceral leishmaniasis cases.

INTRODUÇÃO: A alergia a camarão é causa comum e potencialmente grave de alergia alimentar IgE mediada, incluindo anafilaxia. Ao contrário de outras alergias alimentares, a alergia a camarão afeta predominantemente adultos e geralmente é vitalícia. Até o momento, não existe terapia específica para a alergia a camarão. Os pacientes necessitam excluir o crustáceo de sua dieta e portar adrenalina auto-injetável, para uso em reações por exposição acidental. A complexidade do perfil alergênico do camarão tem sido cada vez mais reconhecida nos últimos dez anos. A proteína muscular tropomiosina foi o primeiro alérgeno do camarão identificado e é considerada o seu principal alérgeno. Alguns estudos indicam que a tropomiosina apresenta alta especificidade na alergia a camarão, sendo apontada como uma possível e importante ferramenta diagnóstica. OBJETIVO: O objetivo deste estudo foi avaliar sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo da tropomiosina em pacientes brasileiros em restrição alimentar por suspeita de alergia a camarão. MÉTODOS: Selecionou-se 32 indivíduos com suspeita de alergia a camarão, os quais foram submetidos a testes cutâneos de leitura imediata para camarão (extrato) e \"in natura\" (cru e cozido), ácaros e barata. Também foram realizadas dosagens séricas de IgE específica para camarão, tropomiosina de camarão, ácaros e barata americana. Avaliou-se reatividade clínica a camarão através de testes de provocação oral. O camarão utilizado nos testes cutâneos e nos testes de provocação oral foi o Xiphopenaeus kroyeri (sete barbas). Foi realizado Western Blot 1 D dos pacientes alérgicos. Foram realizados cálculos de valor de corte para teste cutâneo de leitura imediata e IgE sérica específica através da utilização da curva ROC. RESULTADOS: Alergia a camarão foi confirmada em 17 pacientes. A IgE sérica específica para tropomiosina de camarão neste estudo apresentou sensibilidade de 58,8%, especificidade de 60%, valor preditivo positivo de 62,5% e valor preditivo negativo 56%. Em comparação com a IgE especifica para camarão e os testes cutâneos para camarão com extrato e \"in natura\", a IgE específica para tropomiosina apresentou a menor sensibilidade e menor valor preditivo positivo. Em sete pacientes alérgicos, a tropomiosina não foi detectada, apontando a importância de outros alérgenos do camarão, avaliados no Western blot. Os testes cutâneos apresentaram diferença estatística significativa entre alérgicos e não alérgicos e foi possível definir um ponto de corte, útil na distinção entre ambos. Os resultados dos valores de corte do teste cutâneo (média) para extrato comercial foram 5,75 mm, para camarão cru 6,75 mm e para camarão cozido 5,00 mm. Todos os pacientes são atópicos. CONCLUSÃO: A IgE específica para tropomiosina neste estudo não apresentou superioridade diagnóstica quando comparada aos testes cutâneos com camarão \"in natura\" e extrato, e a IgE específica para o camarão. Outros alérgenos, além da tropomiosina, devem ser considerados na avaliação diagnóstica para a alergia a camarão / INTRODUCTION: Shrimp allergy is a common and potentially serious cause of food allergy-mediated IgE, including anaphylaxis. Unlike other food allergies, shrimp allergy affects predominantly adults and is usually lifelong. To date, there is no specific therapy for shrimp allergy. Patients need to exclude the crustacean from their diet and carry selfinjectable adrenaline to be used if the reaction begins after accidental exposure. The complexity of the allergenic profile of shrimp has been increasingly recognized in the last ten years. The tropomyosin muscle protein was the first identified shrimp allergen and is considered its main allergen. Some studies indicate that tropomyosin presents high specificity in shrimp allergy, being pointed out as a possible and important diagnostic tool. OBJECTIVE: The objective of this study was to evaluate sensitivity, specificity, positive predictive value and negative predictive value of tropomyosin in Brazilian patients under food restriction due to the suspicion of allergy to shrimp. METHODS: Thirty-two individuals with suspected allergy to shrimp were selected, who underwent immediate skin tests for shrimp (extract) and \"in natura\" (raw and cooked), mites and cockroaches. Serum dosages of IgE specific for shrimp, shrimp tropomyosin, mites and American cockroach were also performed. Clinical reactivity to shrimp was assessed by oral challenge tests. Shrimp used in skin tests and oral challenge tests was Xiphopenaeus kroyeri (seabob). Western Blot 1 D of the allergic patients was performed. Cut-off calculations were performed for immediate-reading skin test and specific serum IgE using the ROC curve. RESULTS: Shrimp allergy was confirmed in 17 patients. Serum IgE specific for shrimp tropomyosin in this study showed sensitivity of 58.8%, specificity of 60%, positive predictive value of 62.5% and negative predictive value of 56%. Compared to shrimp-specific IgE and cutaneous shrimp skin tests with extract and \"in natura\", tropomyosin was not detected, indicating the importance of other shrimp allergens evaluated in the Western blot. The skin tests presented a statistically significant difference between allergic and non-allergic and it was possible to define a cutoff point, useful in distinguishing between both. The results of the cutoff values of the skin test (average) for commercial extract were 5.75 mm, for raw shrimp 6.75 mm and for cooked shrimp 5.00 mm. All patients are atopic. CONCLUSION: The tropomyosin-specific IgE in this study did not present diagnostic superiority when compared to cutaneous tests with in natura shrimp and extract, and shrimp specific IgE. Other allergens, in addition to tropomyosin, should be considered in the diagnostic evaluation for shrimp allergy

A fusion protein composed of antibody fragments and β-lactamase was earlier created by Kojima et al. (2011), with antigen specificities against a bone disease marker and a pesticide. The enzyme was circularly permutated and fused to the variable heavy and light chain antibody fragments, thereby ensuring inactivity until binding of the target antigen triggered enzyme activation. Upon activation, the β-lactamase produced a colorimetric signal, which indicated antigen presence. In this work, a similar strategy was used to create two novel fusion proteins composed of circularly permuted β-lactamase and superfolder green fluorescent protein with anti-benzo[a]pyrene variable antibody fragments. The fusion proteins were designed and expressed in E. coli for the development of a single-step visual immunoassay. It was hypothesized that the cp reporter proteins would be activated once the binding of B[a]P to the variable antibody fragments occurred, and this interaction was expected to produce a detectable colorimetric or fluorescent signal. Although positive results were obtained in one instance, substantial supportive evidence in favour of the hypothesis could not be obtained. / SENTINEL Bioactive Paper Network, Natural Sciences and Engineering Research Council of Canada (NSERC), Canada Research Chairs Program.

Made available in DSpace on 2014-07-29T15:07:43Z (GMT). No. of bitstreams: 1 Dissertacao Duanne A da Silva.pdf: 1367420 bytes, checksum: 2450c56c8175888ad1aaa259e17f1059 (MD5) Previous issue date: 2011-02-24 / Herpetic meningoencephalitis is an infection of the central nervous system caused by bovine herpesvirus 5 (BoHV-5) despite BoHV-1 also being associated with this pathology. The course of the disease is usually fatal. Meningoencephalitis has caused significant economic burden to the Brazilian cattle industry due to its high mortality rate, especially in young animals. Because of cross-reactivity and the absence of an effective serological test that differentiates types 1 and 5 of herpesvirus, the actual prevalence of infection with both viruses is unknown. It is estimated that some animals, supposedly infected with type 1, may be seropositive for type 5. For a better understanding of the molecular epidemiology of this virus in Goiás, 110 brain samples of young cattle, that died with neurological signs, referred to the reference laboratory of the State of Goiás for rabies diagnosis in 2008, were analyzed by multiplex PCR to amplify the glycoprotein C (gC) region of the BoHV-1 and -5 DNA. Of the 110 samples, 53.6% were positive for bovine herpesvirus. Of these, 22.0% were positive for BoHV-1, whereas 52.5% were positive for BoHV-5. Furthermore, 13.6% of samples showed co-infection for types 1 and 5. Among these, one sample came from a buffalo. Positivity for both bovine herpesvirus and rabies virus was observed in 53.3% of samples. This study showed that BoHV-1 and -5 are circulating among cattle in the State of Goiás and that herpesvirus type 1 can also be encephalitogenic. Although there was no evidence of viral replication, after inoculation of all 20 brain samples in cell culture, the possibility of viruses being associated with neurological disease cannot be ruled out. This was the first study to demonstrate the presence of BoHV-1 and BoHV-5 in samples obtained from herds in Goiás. / A meningoencefalite herpética é uma infecção do sistema nervoso central de curso geralmente fatal causada pelo herpesvírus bovino tipo 5 (BoHV-5), todavia, também o BoHV-1 pode estar associado a essa patologia. A enfermidade tem gerado muitos prejuízos à pecuária brasileira por apresentar elevada letalidade, principalmente, entre animais jovens. Devido a reatividade cruzada e a falta de um teste sorológico eficaz que diferencie os tipos 1 e 5 de herpesvírus bovino, não se sabe a real prevalência das infecções por ambos os vírus. Estima-se que uma parcela dos animais supostamente infectados pelo tipo 1 possa ser soropositiva para o tipo 5. Para um melhor entendimento da epidemiologia molecular desses vírus em Goiás, 110 amostras de cérebro de bovinos jovens que vieram a óbito com sinais neurológicos, encaminhadas ao laboratório de referência do Estado para diagnóstico de raiva no ano de 2008, foram analisadas por PCR multiplex para amplificação da região da glicoproteína C (gC) do DNA do BoHV-1 e -5. Das 110 amostras, 53,6% foram positivas para herpesvírus bovino. Destas, 22,0% foram positivas para o BoHV-1, enquanto 52,5% foram positivas para o BoHV-5. Além disso, encontrou-se 13,6% de coinfecção para os tipos 1 e 5. Entre estas, uma amostra foi proveniente de um bubalino. Foi observada positividade simultânea para herpesvírus bovino e o vírus da raiva em 53,3% das amostras. Este estudo permitiu concluir que o BoHV-1 e o -5 circulam no Estado de Goiás e que o BoHV-1 apresenta potencial encefalitogênico. Apesar de não ter ocorrido multiplicação viral em nenhuma das 20 amostras de cérebro inoculadas em cultura de célula, não se pode descartar a possibilidade da doença neurológica estar associada aos vírus nas outras amostras analisadas. Este foi o primeiro estudo com amostras de Goiás a comprovar a presença dos dois vírus nos rebanhos do Estado.