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PubMed1990

A model for determining immunogenic relationships between avian infectious bronchitis viruses.

A model for determining immunogenic relationships between strains of infectious bronchitis virus is described that is based upon the vaccinating dose required to induce prevention of multiplication of a standard challenge dose of an homologous strain in the lungs of specific-pathogen-free chickens. The model has been used to demonstrate that: (1) Approximately 100 times the vaccinating dose must be used to produce immunity from one compared with two doses; (2) differences of immunogenicity of greater than 1,000-fold exist between Australian strains; (3) immunogenicity is greatest for live virus administered directly to the trachea, followed by live virus administered by the ocular route, inactivated virus administered intratracheally and inactivated virus administered intramuscularly; (4) for chickens inoculated with Australian vaccine strains, protection against challenge is unrelated to serotype.

Aging↗

PubMedDate not supplied

A rapid-plate hemagglutination assay for the detection of infectious bronchitis virus.

A rapid-plate hemagglutination (HA) test to detect infectious bronchitis virus (IBV) in allantoic fluid of embryonated eggs was introduced into routine procedures for IBV identification. This system was tested in 468 diagnostic cases received by the Poultry Diagnostic and Research Center at the University of Georgia. Allantoic fluids from inoculated embryos were harvested and treated with commercially available neuraminidase enzyme. IBV in neuraminidase-treated allantoic fluid was identified by clear and consistent HA of chicken red blood cells within 1 min of incubation. The specificity of the neuraminidase rapid-plate HA assay was examined with other avian viruses in individual and dual embryonic infections. Sensitivity of this test was compared with embryo lesions and reverse transcriptase-polymerase chain reaction (RT-PCR). The rapid-plate HA assay of neuraminidase-treated allantoic fluid correlated with the RT-PCR during the early stages of IBV detection, identification, and isolation in embryonated eggs.

Allantois↗

PubMed2006

Development and evaluation of a real-time Taqman RT-PCR assay for the detection of infectious bronchitis virus from infected chickens.

It is important to rapidly differentiate infectious bronchitis virus (IBV) from disease agents like highly pathogenic avian influenza virus and exotic Newcastle disease virus, which can be extremely similar in the early stages of their pathogenesis. In this study, we report the development and testing of a real-time RT-PCR assay using a Taqman-labeled probe for early and rapid detection of IBV. The assay amplifies a 143-bp product in the 5'-UTR of the IBV genome and has a limit of detection and quantification of 100 template copies per reaction. All 15 strains of IBV tested as well as two Turkey coronavirus strains were amplified, whereas none of the other pathogens examined, tested positive. Evaluation of the assay was completed with 1329 tracheal swab samples. A total of 680 samples collected from IBV antibody negative birds were negative for IBV by the real-time RT-PCR assay. We tested 229 tracheal swabs submitted to two different diagnostic laboratories and found 79.04% of the tracheal swabs positive for IBV by real-time RT-PCR, whereas only 27.51% of the samples were positive by virus isolation, which is the reference standard test. We also collected a total of 120 tracheal swabs at six different time points from birds experimentally infected with different dosages of IBV and found that, independent of the dose given, the viral load in the trachea plateau at 5 days post-inoculation. In addition, an inverse relationship between the dose of virus given and the viral load at 14 days post-inoculation was observed. Finally, we tested 300 total tracheal swab samples, from a flock of commercial broilers spray vaccinated for IBV in the field. The percentage of birds infected with the IBV vaccine at 3, 7, and 14 days post-vaccination was 58%, 65%, and 83%, respectively, indicating that only slightly more than half the birds were initially infected then the vaccine was subsequently transmitted to other birds in the flock. This observation is significant because coronaviruses, which have a high mutation rate, can revert to pathogenicity when bird-to-bird transmission occurs. The real-time RT-PCR test described herein can be used to rapidly distinguish IBV from other respiratory pathogens, which is important for control of this highly infectious virus. The test was extremely sensitive and specific, and can be used to quantitate viral genomic RNA in clinical samples.

5' Untranslated Regions↗

PubMed1986

Cloning and sequencing of 5' terminal sequences from avian infectious bronchitis virus genomic RNA.

The subgenomic RNAs of the fowl coronavirus infectious bronchitis virus (IBV) form a 3' co-terminal or 'nested' set. The presence of non-contiguous (leader) sequences fused to the 5' termini of murine hepatitis virus mRNAs has been demonstrated using RNase T1 oligonucleotide mapping and sequencing. The presence of a leader sequence on IBV mRNA A has been demonstrated previously. In this paper the presence of a leader identical to that present on the 5' terminus of IBV mRNA A is demonstrated to be present on the 5' terminus of IBV genomic RNA. This has been achieved by sequencing of primer extension products and cDNA clones containing the genomic leader. Analysis of these clones has revealed the presence of a sequence at the leader/genome-length RNA junction which is closely related to regions of homology identified previously within the genomic RNA sequence at the leader/body junctions of subgenomic RNAs. The implications of this finding for mechanisms of coronavirus RNA synthesis are discussed.

Animals↗

PubMed2006

Sialic acid is a receptor determinant for infection of cells by avian Infectious bronchitis virus.

The importance of sialic acid for infection by avian Infectious bronchitis virus (IBV) has been analysed. Neuraminidase treatment rendered Vero, baby hamster kidney and primary chicken kidney cells resistant to infection by the IBV-Beaudette strain. Sialic acid-dependent infection was also observed with strain M41 of IBV, which infects primary chicken kidney cells but not cells from other species. In comparison with Influenza A virus and Sendai virus, IBV was most sensitive to pre-treatment of cells with neuraminidase. This finding suggests that IBV requires a greater amount of sialic acid on the cell surface to initiate an infection compared with the other two viruses. In previous studies, with respect to the haemagglutinating activity of IBV, it has been shown that the virus preferentially recognizes alpha2,3-linked sialic acid. In agreement with this finding, susceptibility to infection by IBV was connected to the expression of alpha2,3-linked sialic acid as indicated by the reactivity with the lectin Maackia amurensis agglutinin. Here, it is discussed that binding to sialic acid may be used by IBV for primary attachment to the cell surface; tighter binding and subsequent fusion between the viral and the cellular membrane may require interaction with a second receptor.

Animals↗

PubMedDate not supplied

Isolation, pathogenicity, and H120 protection efficacy of infectious bronchitis viruses isolated in Taiwan.

Seven isolates of infectious bronchitis (IB) virus (IBV) were isolated from two breeder farms and five broiler farms in Taiwan in 1992. The cardinal signs of disease in breeders were egg production drops and watery albumen, and those in broilers were respiratory distress and renal urate deposition or death. All diseased chickens had been vaccinated with IB vaccines (mostly H120). The viruses were isolated and identified by chicken embryo inoculation and electron microscopy. The genomes of the isolates were extracted and amplified by polymerase chain reaction; the restriction fragment length polymorphism analysis suggested that the genotypes of the present IBV isolates were different from the eight reference strains. One-day-old specific-pathogen-free chicks were inoculated with the field isolates in order to test the virulence of those isolates. Respiratory distress and depression commenced at 24 hours after inoculation. Two of the seven isolates were found to be highly virulent, causing 50% or more mortality in inoculated chicks. Vaccine protection tests showed that H120 could protect chickens against challenges with four of six field isolates.

Animals↗

PubMedDate not supplied

Molecular characterization of an infectious bronchitis virus strain isolated from an outbreak in vaccinated layers.

An infectious bronchitis virus (IBV) strain CT/7852/97 was isolated from a commercial layer flock experiencing decreased egg production and poor egg quality. Reciprocal virus neutralization test demonstrated that the isolate was closely related to Massachusetts, Arkansas, and JMK. With the use of both reverse transcription-polymerase chain reaction-restriction fragment length polymorphism and multiplex polymerase chain reaction methods, results consistently showed that CT/7852/97 isolate was Massachusetts serotype specific only. The nucleotide sequence of S1 polypeptide coding region of CT/7852/97 was determined and compared with the published sequences of 12 IBV strains from North America. The highest nucleotide homology with H120 (97%) and the least homology (62%) with Delaware (De/072/92) were observed. Massachusetts 41 and Beaudette shared 94% and 93%, respectively, nucleotide similarity with CT/7852/97, and Connecticut, JMK, and Arkansas 99 had 94%, 83%, and 78%, respectively, nucleotide similarity with CT/7852/97. Furthermore, no possible recombination site on S1 gene among Massachusetts 41, Arkansas 99, and JMK was detected in the isolate.

Amino Acid Sequence↗

PubMed2000

Identification of a novel cleavage activity of the first papain-like proteinase domain encoded by open reading frame 1a of the coronavirus Avian infectious bronchitis virus and characterization of the cleavage products.

The coronavirus Avian infectious bronchitis virus (IBV) employs polyprotein processing as a strategy to express its gene products. Previously we identified the first cleavage event as proteolysis at the Gly(673)-Gly(674) dipeptide bond mediated by the first papain-like proteinase domain (PLPD-1) to release an 87-kDa mature protein. In this report, we demonstrate a novel cleavage activity of PLPD-1. Expression, deletion, and mutagenesis studies showed that the product encoded between nucleotides 2548 and 8865 was further cleaved by PLPD-1 at the Gly(2265)-Gly(2266) dipeptide bond to release an N-terminal 195-kDa and a C-terminal 41-kDa cleavage product. Characterization of the cleavage activity revealed that the proteinase is active on this scissile bond when expressed in vitro in rabbit reticulocyte lysates and can act on the same substrate in trans when expressed in intact cells. Both the N- and C-terminal cleavage products were detected in virus-infected cells and were found to be physically associated. Glycosidase digestion and site-directed mutagenesis studies of the 41-kDa protein demonstrated that it is modified by N-linked glycosylation at the Asn(2313) residue encoded by nucleotides 7465 to 7467. By using a region-specific antiserum raised against the IBV sequence encoded by nucleotides 8865 to 9786, we also demonstrated that a 33-kDa protein, representing the 3C-like proteinase (3CLP), was specifically immunoprecipitated from the virus-infected cells. Site-directed mutagenesis and expression studies showed that a previously predicted cleavage site (Q(2583)-G(2584)) located within the 41-kDa protein-encoding region was not utilized by 3CLP, supporting the conclusion that the 41-kDa protein is a mature viral product.

3C Viral Proteases↗

PubMed2004

A survey of the prevalence of infectious bronchitis virus type 4/91 in Iran.

To evaluate the prevalence of infectious bronchitis virus (IBV) type 4/91 in Iran, tracheal swabs from 77 broiler flocks in 16 provinces were collected at the slaughterhouse. Swabs were subjected to RNA extraction and tested by RT-PCR, followed by a type-specific nested PCR. The viral RNA was detected in 33 samples (42.8%) from different provinces. The results indicate a relatively high prevalence of IBV type 4/91 in Iran and necessitate revising the vaccination programme against this disease.

Animals↗

PubMed1985

Humoral antibody response and assessment of protection following primary vaccination of chicks with maternally derived antibody against avian infectious bronchitis virus.

Commercially bred chicks with maternally derived antibody to avian infectious bronchitis virus (IBV) were hatched in isolated conditions and a number vaccinated within the first three weeks of life with live IBV strain H120. Humoral antibody responses were assayed by haemagglutination inhibition (HI) or neutralisation (SN) tests, and the degree of protection against challenge with the virulent Massachusetts M41 strain assessed on the basis of tracheal ciliary activity four days after challenge. Maternal antibody in unvaccinated chicks declined linearly with a mean half-life of five to six days based on both HI and SN tests; these chicks were protected against challenge until four weeks old. There was complete correlation between ciliary activity and histopathological findings, but little between protection and antibody titre. It was concluded that the optimum age for primary vaccination was about two weeks.

Animals↗

PubMed2004

Generation of the transgenic potato expressing full-length spike protein of infectious bronchitis virus.

To seek a new delivery system of vaccine for infectious bronchitis virus (IBV), transgenic potato expressing full-length spike (S) protein of IBV was produced and its immunogenicity in chickens was investigated. One to three copies of S gene of IBV were randomly and stably inserted into potato (Solanum tuberrosum cv. Dongnong 303) genome by Agrobacterium tumefaciens-mediated transformation. Transcription and translation of S gene for IBV were confirmed by Northern blot and Western blot analyses in transgenic plantlets. Chickens immunized orally and intramuscularly with transgenic potato tubers expressing S protein generated the detectable levels of serum neutralizing antibodies and were protected against the challenge with the virulent IBV. In vitro secretion of interleukin 2 and T lymphocyte proliferation of spleen cells from the immunized chickens varied with the dose and the manner of vaccination with S protein derived from transgenic plants. The results indicated that S protein expressed in transgenic plants might be a new source for the production of Coronaviridae IBV vaccine.

Animals↗

PubMed1997

Ceramide accumulation uncovers a cycling pathway for the cis-Golgi network marker, infectious bronchitis virus M protein.

The M glycoprotein from the avian coronavirus, infectious bronchitis virus (IBV), contains information for localization to the cis-Golgi network in its first transmembrane domain. We hypothesize that localization to the Golgi complex may depend in part on specific interactions between protein transmembrane domains and membrane lipids. Because the site of sphingolipid synthesis overlaps the localization of IBV M, we asked whether perturbation of sphingolipids affected localization of IBV M. Short-term treatment with two inhibitors of sphingolipid synthesis had no effect on localization of IBV M or other Golgi markers. Thus, ongoing synthesis of these lipids was not required for proper localization. Surprisingly, a third inhibitor, d,l-threo-1-phenyl-2-decanoylamino-3-morpholino- 1-propanol (PDMP), shifted the steady-state distribution of IBV M from the Golgi complex to the ER. This effect was rapid and reversible and was also observed for ERGIC-53 but not for Golgi stack proteins. At the concentration of PDMP used, conversion of ceramide into both glucosylceramide and sphingomyelin was inhibited. Pretreatment with upstream inhibitors partially reversed the effects of PDMP, suggesting that ceramide accumulation mediates the PDMP-induced alterations. Indeed, an increase in cellular ceramide was measured in PDMP-treated cells. We propose that IBV M is at least in part localized by retrieval mechanisms. Further, ceramide accumulation reveals this cycle by upsetting the balance of anterograde and retrograde traffic and/ or disrupting retention by altering bilayer dynamics.

Animals↗

PubMed2026

Molecular characteristics, phylodynamics, and evolutionary changes of avian infectious bronchitis virus detected from chickens in Yunnan Province, 2021-2024.

Avian infectious bronchitis virus (IBV) is endemic in poultry flocks worldwide, posing a significant threat to the global poultry industry. Frequent mixing of free-range local chickens with introduced chickens in Yunnan Province, China, facilitates the transmission, recombination, and mutation of avian IBV, thereby complicating disease prevention and control. In this study, we aimed to investigate the presence of IBV in poultry populations in Yunnan Province. Samples were collected from live poultry markets (LPMs) and breeding farms, comprising 725 randomly sampled cloacal/fecal swabs and 55 tissue samples. IBV-positive samples were confirmed via polymerase chain reaction (PCR), with an overall positivity rate of 0.89% (7/780) for all tested samples. The positivity rate was 0.35% (2/564) in Kunming, 3.7% (2/54) in Zhaotong, 20% (1/5) in Yuxi, and 12.5% (2/16) in Baoshan, while no IBV was detected in samples from Lanping, Xichou, or Ninglang. Six IBV strains, including five GI-19 strains and one GVI-1 strain, were successfully isolated. Phylogenetic analysis further showed that the Yunnan GI-19 strains predominantly clustered with strains originating from Sichuan Province. Sequencing of the S1 gene revealed several amino acids substitutions per isolate in hypervariable regions HVR1-HVR3. Notably, a valine (V) and glycine (G) insertion between amino acid positions 88 and 89 was identified exclusively in isolate F210, a feature rarely reported in IBV. Protein-protein docking analysis indicated that the unique 88-89 insertion in isolate F210 S1 may alter its binding interactions with the host receptor ANPEP. Whole-genome comparison revealed that isolate YX3 shared 97.05% nucleotide identity with strain CK/CH/GX/YL17/2017 from Guangxi, whereas isolates Q47, F13, and F210 shared 96.40%-97.27% identity with strain CK/Henan/H1036/2021 from Henan. Recombination analysis detected obvious recombination events in isolates F13, F210, Q47, and YX3, with GI-22 strains serving as the major parental donors. These genetic characteristics, recombination patterns, and structural insights demonstrate the complex evolutionary dynamics of circulating IBV strains in Yunnan. Continuous molecular epidemiological surveillance combined with functional protein analysis is essential to monitor emerging variants and formulating targeted, effective disease control strategies.

Avian infectious bronchitis virus↗

PubMedDate not supplied

Serotypes of avian infectious bronchitis virus isolates from field cases in Japan.

Eight etiologic agents isolated from field cases in Japan were identified as isolates of infectious bronchitis virus by agar-gel diffusion, buoyant-density determination on sucrose-density centrifugation, and morphological study by electron microscope of the purified viruses. In studies of the antigenic relationships of the eight isolates and six known infectious bronchitis viruses, antigenic diversity of these viruses was recognized from the degrees of relatedness using a plaque reduction in the "constant-virus decreasing-serum" method.

Animals↗

PubMed2000

Leader switching occurs during the rescue of defective RNAs by heterologous strains of the coronavirus infectious bronchitis virus.

A defective RNA (D-RNA), CD-61, derived from the Beaudette strain of the avian coronavirus infectious bronchitis virus (IBV), was rescued (replicated and packaged) using four heterologous strains of IBV as helper virus. Sequence analysis of the genomic RNA from the four heterologous IBV strains (M41, H120, HV10 and D207) identified nucleotide differences of up to 17% within the leader sequence and up to 4.3% within the whole of the adjacent 5' untranslated region (UTR). Analysis of the 5' ends of the rescued D-RNAs showed that the Beaudette leader sequence, present on the initial CD-61, had been replaced with the corresponding leader sequence from the helper IBV strain but the adjacent 5' UTR sequence of the rescued D-RNAs corresponded to the original CD-61 Beaudette sequence. These results demonstrated that the phenomenon of leader switching previously identified for the coronaviruses murine hepatitis virus and bovine coronavirus (BCoV) also occurred during the replication of IBV D-RNAs. Three predicted stem-loop structures were identified within the 5' UTR of IBV. Stem-loop I showed a high degree of covariance amongst the IBV strains providing phylogenetic evidence that this structure exists and is potentially involved in replication, supporting previous observations that a BCoV stem-loop homologue was essential for replication of BCoV defective interfering RNAs.

5' Untranslated Regions↗

PubMed1998

Cloacal inoculation with the Connecticut strain of avian infectious bronchitis virus: an attempt to produce nephropathogenic virus by in vivo passage using cloacal inoculation.

Avian infectious bronchitis virus (IBV) strain Connecticut A-5968 isolated from respiratory tissue of chickens in USA in the 1960s, is considered as representative of respiratory disease causing IBV strains. Specific pathogen free chicks inoculated with the strain via the cloaca replicated the virus more rapidly in their kidneys than did chicks inoculated with the same virus intratracheally. Virus passaged thirteen-times via the cloaca caused stronger nephrotropism and nephropathogenicity than the parent virus. It is suggested that cloacal inoculation of IBV provides new and interesting information concerning the mechanisms of nephropathogenicity of IBV which has became increasingly important worldwide during the last ten years.

Animals↗

PubMed1981

Structural proteins and glycoproteins of infectious bronchitis virus particles labeled during growth in chick embryo cells.

Seven Australian infectious bronchitis viruses incorporated radioactive amino acid label during growth in two types of cell culture. From purified virions, four major polypeptides of molecular weights 91,000 (GP91), 51,000 (VP51), 25,000 (GP25) and 23,000 (VP23) were separated by PAGE. Two minor polypeptides of molecular weights 210,000 (GP210) and 185,000 (GP185) were also identified. 3H-glucosamine was incorporated during growth of virus into GP210, GP185, GP91 and GP25. Bromelain digestion of virions removed GP210, GP185 and GP91, leaving VP51, GP25 and VP23 in a large subviral particle. All seven viruses, which were of six serological subtypes, formed apparently identical polypeptide profiles.

Animals↗

PubMed1998

A common RNA motif in the 3' end of the genomes of astroviruses, avian infectious bronchitis virus and an equine rhinovirus.

In the 3' non-coding region of the genomes of infectious bronchitis virus, an avian coronavirus and the picornavirus equine rhinovirus serotype 2, there is a motif with remarkable similarity, both in sequence and folding, to the second RNA stem-loop from the 3' end of the genomes of human astroviruses. This motif was also found in astroviruses of sheep, pig and turkey, suggesting that it is a common feature of all astroviruses. The conserved nature of the motif indicates that there has been strong selection for its preservation. There is significant homology between the regions flanking this motif in infectious bronchitis virus and a continuous RNA sequence at the same distance from the 3' poly(A) tail in some related mammalian coronaviruses. These observations suggest that the presence of the motif in these three viral families is the result of at least two separate RNA recombination events.

Animals↗