scholarly journals Carbonized Lysine-Nanogels Protect against Infectious Bronchitis Virus

2021 ◽  
Vol 22 (11) ◽  
pp. 5415
Author(s):  
Ding-Li Chou ◽  
Ju-Yi Mao ◽  
Anisha Anand ◽  
Han-Jia Lin ◽  
John Han-You Lin ◽  
...  
Keyword(s):  
Infectious Bronchitis Virus ◽  
Pseudorabies Virus ◽  
Hatching Rate ◽  
Viral Inhibition ◽  
Infected Chicken ◽  
In Ovo ◽  
Chicken Embryos ◽  
Virus Attachment ◽  
Infectious Bronchitis

In this study, we demonstrate the synthesis of carbonized nanogels (CNGs) from an amino acid (lysine hydrochloride) using a simple pyrolysis method, resulting in effective viral inhibition properties against infectious bronchitis virus (IBV). The viral inhibition of CNGs was studied using both in vitro (bovine ephemeral fever virus (BEFV) and pseudorabies virus (PRV)) and in ovo (IBV) models, which indicated that the CNGs were able to prevent virus attachment on the cell membrane and penetration into the cell. A very low concentration of 30 μg mL−1 was found to be effective (>98% inhibition) in IBV-infected chicken embryos. The hatching rate and pathology of IBV-infected chicken embryos were greatly improved in the presence of CNGs. CNGs with distinctive virus-neutralizing activities show great potential as a virostatic agent to prevent the spread of avian viruses and to alleviate the pathology of infected avian species.

scholarly journals Treatment with Exogenous Trypsin Expands In Vitro Cellular Tropism of the Avian Coronavirus Infectious Bronchitis Virus

Viruses ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 1102
Author(s):  
Phoebe Stevenson-Leggett ◽  
Sarah Keep ◽  
Erica Bickerton
Keyword(s):  
Cell Culture ◽  
Infectious Bronchitis Virus ◽  
Ex Vivo ◽  
Serial Passage ◽  
In Ovo ◽  
Infectious Bronchitis ◽  
Protease Treatment ◽  
Cellular Tropism ◽  
Passage Cell

The Gammacoronavirus infectious bronchitis virus (IBV) causes a highly contagious and economically important respiratory disease in poultry. In the laboratory, most IBV strains are restricted to replication in ex vivo organ cultures or in ovo and do not replicate in cell culture, making the study of their basic virology difficult. Entry of IBV into cells is facilitated by the large glycoprotein on the surface of the virion, the spike (S) protein, comprised of S1 and S2 subunits. Previous research showed that the S2′ cleavage site is responsible for the extended tropism of the IBV Beaudette strain. This study aims to investigate whether protease treatment can extend the tropism of other IBV strains. Here we demonstrate that the addition of exogenous trypsin during IBV propagation in cell culture results in significantly increased viral titres. Using a panel of IBV strains, exhibiting varied tropisms, the effects of spike cleavage on entry and replication were assessed by serial passage cell culture in the presence of trypsin. Replication could be maintained over serial passages, indicating that the addition of exogenous protease is sufficient to overcome the barrier to infection. Mutations were identified in both S1 and S2 subunits following serial passage in cell culture. This work provides a proof of concept that exogenous proteases can remove the barrier to IBV replication in otherwise non-permissive cells, providing a platform for further study of elusive field strains and enabling sustainable vaccine production in vitro.

REDUCED CHICKEN EMBRYO DWARFING EFFECT IS RELATED TO INFECTIOUS BRONCHITIS VIRUS TW2575/98 REPLICATION EFFICIENCY

2020 ◽  
Vol 46 (02n03) ◽  
pp. 85-93
Author(s):  
Cheng-Ta Tsai ◽  
Ming-Chang Lee ◽  
Ching-Ho Wang
Keyword(s):  
Vaccine Strain ◽  
Infectious Bronchitis Virus ◽  
Tissue Tropism ◽  
Renal Tubules ◽  
Pathological Changes ◽  
In Ovo ◽  
Chicken Embryos ◽  
Infectious Bronchitis ◽  
Attenuated Vaccine ◽  
Embryonic Death

An attenuated infectious bronchitis virus (TW2575/98) vaccine strain was successfully developed after 75 serial passages in embryonated chicken eggs. However, the in ovo vaccination for disease control was not applied in practice because this vaccine strain is highly pathogenic to chicken embryos (CEs) causing early death, dwarfing and other harmful effects. We compared the differences in virus replication, pathological changes, and tissue tropism between the wild virus and attenuated vaccine strain in CEs inoculated with different viral titer levels, i.e. 0.1, 1 and 10 EID[Formula: see text]/egg. The wild virus caused dwarfing effect at high titer inoculation, whereas the attenuated vaccine strain caused the dwarfing effect only at a lower viral inoculation accompanied by the earlier infection establishment and embryonic death at high and medium titers. There were no significant differences in the pathological changes in CEs infected by both wild and attenuated strains. Detected by immunohistochemistry, the viral antigens of both strains could be found mainly at the epithelium of the chorioallantoic membrane, lung parabronchus, renal tubules and some in the spleen and heart serosa. These findings indicated that the early embryonic death and dwarfing is not related to the change in cell/tissue tropism of the vaccine strain, rather on the early infection establishment and viral load. We suggest that the vaccine strain inoculated titer could be adjusted to an optimal low level for in ovo vaccination to overcome the poor hatching rate for its higher virulence to chicken embryos.

scholarly journals In Vitro and In Ovo Expression of Chicken Gamma Interferon by a Defective RNA of Avian Coronavirus Infectious Bronchitis Virus

2003 ◽  
Vol 77 (10) ◽  
pp. 5694-5702 ◽  
Author(s):  
Karen Hackney ◽  
Dave Cavanagh ◽  
Pete Kaiser ◽  
Paul Britton
Keyword(s):  
Infectious Bronchitis Virus ◽  
Allantoic Fluid ◽  
Helper Virus ◽  
Gamma Interferon ◽  
Biologically Active ◽  
Site Directed Mutagenesis ◽  
In Ovo ◽  
Infectious Bronchitis

ABSTRACT Coronavirus defective RNAs (D-RNAs) have been used for site-directed mutagenesis of coronavirus genomes and for expression of heterologous genes. D-RNA CD-61 derived from the avian coronavirus infectious bronchitis virus (IBV) was used as an RNA vector for the expression of chicken gamma interferon (chIFN-γ). D-RNAs expressing chIFN-γ were shown to be capable of rescue, replication, and packaging into virions in a helper virus-dependent system following electroporation of in vitro-derived T7 RNA transcripts into IBV-infected cells. Secreted chIFN-γ, under the control of an IBV transcription-associated sequence derived from gene 5 of the Beaudette strain, was expressed from two different positions within CD-61 and shown to be biologically active. In addition, following infection of 10-day-old chicken embryos with IBV containing D-RNAs expressing chIFN-γ, the allantoic fluid was shown to contain biologically active chIFN-γ, demonstrating that IBV D-RNAs can express heterologous genes in vivo.

scholarly journals Tissue Distribution of Avian Infectious Bronchitis Virus following in Ovo Inoculation of Chicken Embryos Examined by in Situ Hybridization with Antisense Digoxigenin-Labeled Universal Riboprobe

2002 ◽  
Vol 14 (5) ◽  
pp. 377-381 ◽  
Author(s):  
Chang-Won Lee ◽  
Corrie Brown ◽  
Mark W. Jackwood
Keyword(s):  
In Situ Hybridization ◽  
Infectious Bronchitis Virus ◽  
Tissue Tropism ◽  
Positive Staining ◽  
In Ovo ◽  
Chicken Embryos ◽  
Infectious Bronchitis ◽  
Different Strains

Chicken embryos were inoculated with 8 different strains of infectious bronchitis virus (IBV) representing 7 different serotypes at 17 days of embryonation. At 2 and 5 days postinfection (dpi), tissues were collected for in situ hybridization using an antisense digoxigenin-labeled riboprobe corresponding to the sequence of the mRNA coding for the membrane protein. Extensive antigen staining in the cytoplasm of epithelial cells in the trachea, lung, bursa, and intestine was detected at 2 dpi with all 8 strains of IBV. At 5 dpi, little or no positive staining was observed in these tissues. However, tubular cells of the kidney showed multifocal positive staining with the Wolgemuth strain-, Gray strain-, JMK strain-, and Mass41 strain-infected chickens. No viral RNA was detected in the spleen at any time point. The results demonstrated strict epitheliotropic nature and wide tissue tropism of strains of IBV in the chicken embryo and the universality of our riboprobe. In situ hybridization with this probe will be useful for understanding the tissue tropism and the pathogenesis of IBV in vivo.

Comparative Evaluation of In Vitro and In Vivo Assays for the Detection of Avian Infectious Bronchitis Virus as a Contaminant of Live Poultry Vaccines

1998 ◽  
Vol 26 (5) ◽  
pp. 629-634
Author(s):  
Emiliana Falcone ◽  
Edoardo Vignolo ◽  
Livia Di Trani ◽  
Simona Puzelli ◽  
Maria Tollis
Keyword(s):  
Infectious Bronchitis Virus ◽  
Serological Response ◽  
Avian Infectious Bronchitis Virus ◽  
Animal Testing ◽  
Rt Pcr ◽  
Pcr Assay ◽  
In Vivo Test ◽  
Infectious Bronchitis

A reverse transcriptase polymerase chain reaction (RT-PCR) assay specific for identifying avian infectious bronchitis virus (IBV) in poultry vaccines, and the serological response to IBV induced by the inoculation of chicks with a Newcastle disease vaccine spiked with the Massachusetts strain of IBV, were compared for their ability to detect IBV as a contaminant of avian vaccines. The sensitivity of the IBV-RT-PCR assay provided results which were at least equivalent to the biological effect produced by the inoculation of chicks, allowing this assay to be considered a valid alternative to animal testing in the quality control of avian immunologicals. This procedure can easily be adapted to detect a number of contaminants for which the in vivo test still represents the only available method of detection.

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