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1. Comparative Study of Three Different ELISA to Measure the Antibodies Against Infectious Bronchitis Virus in Vaccinated and Unvaccinated Broilers

Author

Cardoso, TC [UNESP], Oliveira, C, Da-Silva, SEL [UNESP], Ferreira, HL [UNESP], Pinto, AA [UNESP], and Universidade Estadual Paulista (Unesp)

Subjects
infectious bronchitis virus, broilers, post vaccinal response, ELISA, resposta pós vacinal, frangos, vírus da bronquite infecciosa
Abstract

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T19:08:06Z No. of bitstreams: 0 Made available in DSpace on 2013-08-22T19:08:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2001-01-01 Made available in DSpace on 2013-09-30T20:13:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2001-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:18:26Z No. of bitstreams: 0 Made available in DSpace on 2014-05-20T15:18:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2001-01-01 No presente estudo, foram vacinados 150 frangos de corte de um dia de idade com sorotipo H120, e, após 28 dias desafiados à vacinação com o sorotipo M 41 do vírus da bronquite infecciosa das aves. da mesma forma, foram obtidos 150 soros de aves não vacinadas para a análise. Os respectivos soros foram analisados 28, 34 e 46 dias após o desafio, examinados através das técnicas de ELISA indireto (ELISA-I), Sandwich ELISA (S-ELISA) e ELISA com bloqueio de fase líquida (ELISA-BFL) e comparados com a técnica padrão de soroneutralização (SN) para efeito de cálculo da especificidade e sensibilidade relativas, bem como os valores predictivos positivos e negativos. O cálculo do coeficiente de correlação também foi empregado para a análise de concordância. Assim, os valores da correlação encontrados foram r = 0.98 entre ELISA-BFL x SNT, r = 0.79 entre S-ELISA x SNT e r = 0.74 ELISA-I x SNT. No entanto, quando comparamos as técnicas de ELISA entre si, ELISA-BFL x S-ELISA, ELISA-BFL x ELISA-I e S-ELISA x ELISA-I os valores encontrados foram r = 0.75, r = 0.69 e r = 0.79. A técnica de ELISA-BFL demonstrou melhor sensibilidade relativa que as técnicas de S-ELISA e ELISA-I, mesmo 46 dias após o desafio com a estirpe heteróloga. Entretanto, apesar das técnicas de S-ELISA e ELISA-I apresentarem especificidade realtiva superiores, a melhor correlação observada foi entre as técnicas de ELISA-BFL e a SN. Broilers were spray-vaccinated (n=150) with H120 serotype at one-day-old, challenged after 28 days with M41 IBV serotype and after bled at day 28, 34 and 46 after challenged. The respective sera were tested by the indirect ELISA (I-ELISA), sandwich ELISA (S-ELISA), liquid phase blocking ELISA (LPB-ELISA) and the standard serum neutralization test (SNT). For this purpose, a total of 300 sera samples, 150 from non vaccinated and 150 from vaccinated broilers were titrated by all the serological methods and the correlation coefficients were determined. The correlation values (r) between LPB-ELISA x SNT, S-ELISA x SNT, I-ELISA x SNT found werer = 0.98, r = 0.79, and r = 0.74, respectively. Nevertheless, the r between LPB-ELISA x S-ELISA, LPB-ELISA x I-ELISA and S-ELISA x I-ELISA were r = 0.75, r = 0.69 and r = 0.79. In fact , the I-ELISA and the S-ELISA had almost the same correlation with S-ELISA and LPB-ELISA, in contrast with the I-ELISA and LPB-ELISA. It was concluded that LPB-ELISA showed better sensitivity than I-ELISA and S-ELISA, also after 46 days challenge with the heterologous serotype. However, these two last techniques demonstrated similar specificity when the titers were compared with those obtained in SNT, even though after the heterologous serotype challenge the SNT produced better results. This study demonstrated the close relationship between LPB-ELISA and SNT assays. UNESP, Laboratório Multiusuário de Imunodiagnóstico Faculdade de Odontologia Produção e Saúde Animal UNESP Laboratório de Imunologia e Virologia FCAV UNESP, Laboratório Multiusuário de Imunodiagnóstico Faculdade de Odontologia Produção e Saúde Animal UNESP Laboratório de Imunologia e Virologia FCAV

Published
2001

4. Retraction Note to: Mitochondrial bioenergy alterations in avian HD11 macrophages infected with infectious bronchitis virus.

Author

da Silva SEL, Ferreira HL, Garcia AF, Silva FES, Gameiro R, Fabri CUF, Vieira DS, and Cardoso TC

Abstract

The Editor-in-Chief has retracted this article [1]. Figures 1A, 1D and 2B (bottom right) are identical with Figures 1A, 1H and 1B respectively in another article [2] which reports a study in a different species. In addition, Table 1 contains data presented in a third article [3], which also reports a study in a different species. The Editor-in-Chief therefore no longer has confidence in the validity of the data and the conclusions drawn. Tereza C. Cardoso disagrees with this retraction. Helena L. Ferreira agrees with this retraction. Sergio E. L. da Silva, Andrea F. Garcia, Felipe E. S. Silva, Roberto Gameiro, Carolina U. F. Fabri and Dielson S. Vieira have not responded to any correspondence about this retraction.

Published
2020
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5. Mitochondrial bioenergy alterations in avian HD11 macrophages infected with infectious bronchitis virus.

Author

da Silva SEL, Ferreira HL, Garcia AF, Silva FES, Gameiro R, Fabri CUF, Vieira DS, and Cardoso TC

Subjects
Animals, Apoptosis, Cell Line, Cell Proliferation, Chickens, Infectious bronchitis virus growth & development, Macrophages metabolism, Membrane Potential, Mitochondrial, Mitochondria metabolism, Reactive Oxygen Species metabolism, Virion growth & development, Virus Replication, Host-Pathogen Interactions, Infectious bronchitis virus pathogenicity, Macrophages virology, Mitochondria virology, Virion pathogenicity
Abstract

To establish an association between mitochondrial dysfunction and apoptosis following infectious bronchitis virus (IBV) infection, HD11 avian macrophage cells were infected with the Massachusetts 41 (M41) strain. Our results show that the M41 strain of IBV induced cytopathic effects followed by the release of new viral particles. Elevated numbers of apoptotic cells were observed at 24, 48 and 72 h post-infection (p.i.). Viral infection was associated with mitochondrial membrane depolarization and reactive oxygen species (ROS) production at all of the examined timepoints p.i. In summary, IBV M41 replication in infected HD11 macrophages seems to induce mitochondrial bioenergy failure, acting as a respiratory chain uncoupler, without compromising viral replication.

Published
2018
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6. Expression of miR-155 associated with Toll-like receptors 3, 7, and 9 transcription in the olfactory bulbs of cattle naturally infected with BHV5.

Author

Oliveira BRSM, Vieira FV, de S Vieira D, da Silva SEL, Gameiro R, Flores EF, and Cardoso TC

Subjects
Animals, Cattle, Female, Gene Expression Regulation, Herpesvirus 5, Bovine, Olfactory Bulb metabolism, Olfactory Receptor Neurons metabolism, Toll-Like Receptor 3 biosynthesis, Toll-Like Receptor 7 biosynthesis, Toll-Like Receptor 9 biosynthesis, Transcription, Genetic, Encephalitis, Viral metabolism, Herpesviridae Infections metabolism, Meningoencephalitis metabolism, MicroRNAs metabolism, Toll-Like Receptors biosynthesis
Abstract

Bovine herpesvirus 5 (BHV5) infection of young cattle is frequently associated with fatal neurological disease and, as such, represents an attractive model for studying the pathogenesis of viral-induced meningoencephalitis. Following replication in the nasal mucosa, BHV5 invades the central nervous system (CNS) mainly through the olfactory pathway. The innate immune response triggered by the host face to virus replication through the olfactory route is poorly understood. Recently, an upregulation of conserved pathogen-associated molecular pattern, as Toll-like receptors (TLRs), has been demonstrated in the CNS of BHV5 experimentally infected cows. A new perspective to understand host-pathogen interactions has emerged elucidating microRNAs (miRNAs) network that interact with innate immune response during neurotropic viral infections. In this study, we demonstrated a link between the expression of TLRs 3, 7, and 9 and miR-155 transcription in the olfactory bulbs (OB) of 16 cows suffering from acute BHV5-induced neurological disease. The OBs were analyzed for viral antigens and genome, miR-155 and TLR 3, 7, and 9 expression considering three major regions: olfactory receptor neurons (ORNs), glomerular layer (GL), and mitral cell layer (ML). BHV5 antigens and viral genomes, corresponding to glycol-C gene, were detected in all OBs regions by fluorescent antibody assay (FA) and PCR, respectively. TLR 3, 7, and 9 transcripts were upregulated in ORNs and ML, yet only ORN layers revealed a positive correlation between TLR3 and miR-155 transcription. In ML, miR-155 correlated positively with all TLRs studied. Herein, our results evidence miR-155 transcription in BHV5 infected OB tissue associated to TLRs expression specifically ORNs which may be a new window for further studies.

Published
2017
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