Infectious Bronchitis Virus Vaccine Research Articles

Genetic and Antigenic Diversity of Infectious Bronchitis Virus in South America.

Infectious bronchitis virus (IBV) is one of the economically most important diseases affecting the South American poultry industry. The extensive genomic heterogeneity of IBV is a consequence of high mutation rates and recombination events followed by selection. Nucleotide heterogeneity is much higher in the S1 coding region of the relevant spike protein; thus, the S1 sequence is widely used for the IBV genetic classification in genotypes and lineages. Two main lineages (GI-11 and GI-16) extensively circulate in South American chicken flocks. The GI-11 lineage, found exclusively in South America, emerged in the 1950s and is currently the predominant lineage in Brazil and Uruguay. The GI-16 lineage emerged around 1979 and is now circulating in most South American regions. All South American countries include Massachusetts-type strains (GI-1 lineage) in the IBV vaccination programs. The GI-11 and GI-16 lineages display very low antigenic relatedness to Massachusetts vaccine strains. Because these vaccine strains may not confer complete protection against South American lineages, other vaccination strategies have been reported to control GI-11 and GI-16 outbreaks. Analysis of the few full-length genomes of South American strains highlights a complex recombination history of IBV in the continent. A broader geographic and temporal sampling is needed to understand the pattern of genetic variability and the evolutionary history of IBV variants in South America.

Route of infectious bronchitis virus vaccination determines the type and magnitude of immune responses in table egg laying hens

Chicken immune responses to infectious bronchitis virus (IBV) vaccination can depend on route of administration, vaccine strain and bird age. Typically for layer chickens, IBV vaccinations are administered by spray in the hatchery at day-old and boosted at intervals with live vaccines via drinking water (DW). Knowledge of live attenuated IBV vaccine virus kinetics and the immune response in egg-laying hens is exceptionally limited. Here, we demonstrated dissemination of vaccine viruses and differences in hen innate, mucosal, cellular and humoral immune responses following vaccination with Massachusetts or 793B strains, administered by DW or oculonasal (ON) routes. Detection of IBV in the Mass-vaccinated groups was greater during early time-points, however, 793B was detected more frequently at later timepoints. Viral RNA loads in the Harderian gland and turbinate tissues were significantly higher for ON-Mass compared to all other vaccinated groups. Lachrymal fluid IgY levels were significantly greater than the control at 14 days post-vaccination (dpv) for both vaccine serotypes, and IgA mRNA levels were significantly greater in ON-vaccinated groups compared to DW-vaccinated groups, demonstrating robust mucosal immune responses. Cell mediated immune gene transcripts (CD8-α and CD8-β) were up-regulated in turbinate and trachea tissues. For both vaccines, dissemination and vaccine virus clearance was slower when given by DW compared to the ON route. For ON administration, both vaccines induced comparable levels of mucosal immunity. The Mass vaccine induced cellular immunity to similar levels regardless of vaccination method. When given either by ON or DW, 793B vaccination induced significantly higher levels of humoral immunity.

Metagenomic sequencing determines complete infectious bronchitis virus (avian Gammacoronavirus) vaccine strain genomes and associated viromes in chicken clinical samples.

Infectious bronchitis virus (IBV, genus Gammacoronavirus) causes an economically important and highly contagious disease in chicken. Random primed RNA sequencing was applied to two IBV positive clinical samples and one in ovo-passaged virus. The virome of a cloacal swab pool was dominated by IBV (82% of viral reads) allowing de novo assembly of a GI-13 lineage complete genome with 99.95% nucleotide identity to vaccine strain 793B. In addition, substantial read counts (16% of viral reads) allowed the assembly of a near-complete chicken astrovirus genome, while lower read counts identified the presence of chicken calicivirus and avian leucosis virus. Viral reads in a respiratory/intestinal tissue pool were distributed between IBV (22.53%), Sicinivirus (Picornaviridae, 24%), and avian leucosis virus (37.04%). A complete IBV genome with 99.95% nucleotide identity to vaccine strain H120 (lineage GI-1), as well as a near-complete avian leucosis virus genome and a partial Sicinivirus genome were assembled from the tissue sample data. Lower read counts identified chicken calicivirus, Avibirnavirus (infectious bursal disease virus, assembling to 98.85% of segment A and 69.66% of segment B closely related to D3976/1 from Germany, 2017) and avian orthoreovirus, while three avian orthoavulavirus 1 reads confirmed prior real-time RT-PCR result. IBV sequence variation analysis identified both fixed and minor frequency variations in the tissue sample compared to its in ovo-passaged virus. Metagenomic methods allow the determination of complete coronavirus genomes from clinical chicken samples while providing additional insights in RNA virus sequence diversity and coinfecting viruses potentially contributing to pathogenicity.Supplementary InformationThe online version contains supplementary material available at 10.1007/s11262-021-01872-7.

Research Note: Effects of Escherichia coli co-infection on the protective efficacy assessment of two common infectious bronchitis vaccines

Avian infectious bronchitis (IB), a highly contagious disease hazardous to the poultry industry, is caused by an etiological agent called the infectious bronchitis virus (IBV). Some IBV strains (IBVs) alone usually do not cause high mortality in field conditions if not with secondary pathogens including Escherichia coli (E. coli). Herein, we established an IBV and E. coli co-infection model to evaluate the protective efficacy of two IBV vaccine strains against a new emerging genotype GVI-1 with mild virulence in experimental conditions. Chickens were inoculated with IBV field isolate ZQX (genotype GVI-1) and challenged 4 dlater with the E. coli strain MS160427 (serotype O8). Subsequently, these chickens were euthanized at seven days postchallenge (d.p.c.) with E. coli. An autopsy revealed that lesions in the IBV plus E. coli co-infection group were more severe than those in the IBV-infected group. This pathological model was used to assess the protective effect of two commonly used vaccine strains (H120 and 4/91) against the IBV ZQX strain, and a significantly better protective efficacy was observed for 4/91 compared with H120. Thus, IBV and E. coli co-infection could be employed in assessing the protective efficacy of IBV vaccines.

Characterization of an Infectious Bronchitis Virus Isolated from Commercial Layers Suffering from False Layer Syndrome.

Infectious bronchitis virus (IBV) is a gammacoronavirus that primarily induces an upper respiratory disease in chickens, also affecting the urogenital tract and occasionally leading to a condition called false layer syndrome (FLS), where sexually mature hens ovulate normally but are unable to lay eggs. Here, we describe an outbreak of FLS in Arizona from which an IBV variant that is nearly 90% homologous to DMV/1639 using the Spike subunit 1 gene, named AZ/FLS/17, was isolated and used in challenge experiments. Three-day-old specific-pathogen-free chicks were challenged with AZ/FLS/17 or M41 in high and low doses, and the disease outcomes were compared. Overall, no differences in microscopic lesions or viral loads in the reproductive tract were detected between AZ/FLS/17- and M41-infected birds. To minimize the losses linked to FLS in the problematic flocks, an updated live-attenuated IBV vaccine protocol including the use of the Ma5 strain at the hatchery was implemented, resulting in a drastic reduction of false layers in the subsequent flocks. To monitor the circulation of wild-type and vaccine strains in this population, a molecular surveillance study was performed. Samples were collected at 1, 7, 14, and 21 days of age, and from laying hens at 30 and 36 wk. In older birds, the IBV strains detected were more diverse than at 1 and 7 days of age. Nevertheless, live vaccine combinations are still widely used to decrease the losses caused by FLS in commercial egg laying flocks worldwide.

Immunogenicity and efficacy of a bivalent vaccine against infectious bronchitis virus

Infectious bronchitis (IB) is a highly contagious viral disease and is responsible for considerable economic losses in the poultry industry, worldwide. To mitigate the IB-associated losses, multiple vaccines are being applied in the sector with variable successes and thus necessitating the development of a potent vaccine to protect against the IB in the poultry. In the present study, we investigated a bivalent live attenuated vaccine consisting of IB virus (IBV) strain H120 (GI-1 lineage) and D274 (GI-12 lineage) to evaluate its protection against heterologous variant of IBV (GI-23 lineage) in chicken. Protection efficacy was evaluated based on the serology, clinical signs, survival rates, tracheal and kidney histopathology and the viral shedding. Results demonstrated that administering live H120 and D274 (named here Classivar®) vaccine in one day-old and 14 days-old provided 100 % protection. We observed a significant increase in the mean antibody titers, reduced virus shedding, and ameliorated histopathology lesions compared to routinely used vaccination regimes. These results revealed that usage of different IBV vaccines combination can successfully ameliorate the clinical outcome and pathology in vaccinated chicks especially after booster vaccination regime using Classivar®. In conclusions, our data indicate that Classivar® vaccine is safe in chicks and may serve as an effective vaccine against the threat posed by commonly circulating IBV strains in the poultry industry.

The experimental infection with a field isolate of the infectious bronchitis virus from eastern Saudi Arabia resulted in seroconversion of the challenged birds with no apparent clinical diseases.

The infectious bronchitis virus (IBV) is still one of the major respiratory viral pathogens of chickens. The IBV infection resulted in a wide range of clinical syndromes in the affected chickens, including respiratory, renal, gonads affections as well as generalized infections. Despite the intensive application of various commercial vaccines against the virus, many outbreaks are still reported in chickens worldwide. Several studies reported the circulation of several strains and genotypes of the IBV in eastern Saudi Arabia. The main goal of the current study was to isolate some of the circulating strains of IBV and assess its ability to reproduce the IBV infections in the challenge birds. Another objective was to monitor the immune status of the infected chickens during the course of this study. To achieve these goals, we used some filed IBV isolates retrieved from an outbreak in a broiler chicken farm in eastern Saudi Arabia in 2014. A total of 220-day-old chickens (110 Ross and 110 native Saudi breed chickens), twenty birds per each group, were used in this study. The chickens in some groups received some IBV vaccines on day one of the experiment, and some are boosted on day 19. All birds were challenged on day 28 of the experiment. Our results showed mild IBV signs in the non-vaccinated control group of chickens; however, the vaccinated chickens did not show any signs of IBV infections. Meanwhile, both the vaccinated and the none- vaccinated birds seroconverted to the IBV as shown by the ELISA results. In conclusion, the response of the IBV infected birds is mainly driven by the vaccination plans they received as a prime-boost regime. Further studies are required for a better understanding of the dynamics of IBV infection in native Saudi chickens.

Protective Efficacy of Different Live Attenuated Infectious Bronchitis Virus Vaccine Combination against Challenge with GI11 (BR-Type) Strain

The infectious bronchitis virus (IBV) is an avian coronavirus that causes a highly contagious disease that results in substantial economic losses to the poultry industry worldwide. The prevention is mostly based on biosecurity measures and vaccination. The vaccination programs are defined by epidemiological status and cross-immunity provided against different serotypes. Programs using vaccine combinations can be designed to increase protection. We conducted an experimental study to compare two vaccine programs on protection against a virulent strain belonging to the GI-11 genotype (BR-type) isolated in Brazil using two different Mass-type vaccines combined with a BR-type live vaccine. Two groups of commercial chicks were vaccinated at day-1 using two commercially available Mass-type vaccines + BR-type vaccine. They were challenged intranasally at 28-day using 104 EID50/0.1 microliter/chick of wild-type G11 IBV (IBV/24W). Ciliostasis, macroscopic and microscopic lesion scores were evaluated at 4 and 11-days post vaccination (dpv) and 5 days post infection (dpi) and not showed differences between vaccinated groups. MLV+BR-Type group presented highest amount of BR strain vaccine and lowest amount of Mass strain vaccine in tracheal and cloacal swabs at 4 and 11 dpv. In the other hand, the Ma5+BR-type group showed fewer BR-type positive in both samples tested. After challenge, MLV+BR group showed higher titer of antibody measured by ELISA. These results suggest that the selection of a Mass-type vaccine strain can impact directly in the BR vaccine replication, when used associated, and consequently the induction of humoral immune response. Despite that, both protocols provided protection against the challenge with G11 strain.

Effect of dietary stevia-based sweetener on body weight and humoral immune response of broiler chickens.

Background and Aim:Steviol glycosides extracted from the leaves of Stevia rebaudiana Bertoni have been of much consideration recently because of their beneficial effects on health, raising the possibilities for improving farm animals’ health. Although some studies on stevia’s dietary effect on body weight gain are available, few studies have been conducted to evaluate stevioside supplementation on immune response in broilers. This experiment aimed to analyze how a stevia-based sweetener can affect broiler chickens’ growth performance and humoral response.Materials and Methods:In this experiment, one hundred and twenty 1-day-old Cobb-line broiler chicks fed with commercial starter/grower diets were included in three groups and supplemented with stevia-based sweetener at levels 0, 80, and 160 ppm, respectively. Chickens were weighed on day 0 and every 7 days for the next 6 weeks. Chicks were then immunized on days 10 and 24 with a Newcastle and infectious bronchitis vaccine and blood sampled on days 7, 24, and 35. Serologic assays were performed to detect specific antibody levels.Results:The body weight means and body weight gain on day 42 were found to be significantly higher in birds from the group fed with 80 ppm of stevia-based sweetener than those in the control group and slightly higher than those in the group supplemented with 160 ppm of stevia-based sweetener. Likewise, on day 35, antibodies against the Newcastle disease virus were higher in the treatment groups. Immune response to infectious bronchitis virus vaccination was not statistically different among the three groups through the experiment.Conclusion:Stevia-based sweetener at 80 ppm in commercial-based diets improved body weight gain and immune response in broiler chickens at the market age.

A Peptide-Based Enzyme-Linked Immunosorbent Assay for Detecting Antibodies Against Avian Infectious Bronchitis Virus.

Infectious bronchitis virus (IBV) causes substantial loss to the poultry industry despite extensive vaccination. Assessing the antibody response is important for the development and evaluation of effective vaccines. We have developed an enzyme-linked immunosorbent assay (ELISA) for the detection of IBV-specific antibodies, using a synthetic peptide based on a conserved sequence in the IBV spike protein. This peptide-based ELISA (pELISA) specifically detects antibodies to different genotypes of IBV but not antibodies against other common chicken viruses. This assay could detect IBV-specific antibody response on as early as day 7 postinfection. In the testing with field serum samples collected from chickens administered with IBV vaccines, the sensitivity, specificity, and accuracy of pELISA were 98.30, 94.12, and 98.8%, respectively, relative to indirect immunofluorescence assay. Our data demonstrate that the pELISA is of value for the detection of IBV antibody and the evaluation of IBV vaccines.

The efficacy of infectious bronchitis virus vaccination in Central Anatolia

Özet: Enfeksiyöz bronşit virüsü (IBV) kümes hayvanlarında solunum sistemini etkileyen, akut ve hızla yayılan, yumurta verim ve kalitesini etkileyerek ekonomik kayıplara neden olan bir virüstür. Dünya çapında bir dağılımı ve % 100'e ulaşabilen yüksek morbidite oranı vardır. Araştırmamızda İç Anadolu Bölgesi’ndeki 11 yumurtacı kümesten alınan, 599 adet kan örneği incelenmiştir. IBV ile aşılanan kümeslerden toplanan 599 adet kan serum örneğinde, bir kümeste, düşük titre (842) ve yüksek CV (% 84) saptanmıştır. Tüm sonuçlar değerlendirildiğin de iki kümeste ise antikor titresi 2318 ve 1283 elde edilirken, %CV değerleri sırasıyla 93 ve 90 bulunmuştur. Diğer 8 kümesten elde edilen antikor titrelerinin 1 ile 18903, % CV değerlerinin ise 0 ile 63 arasında değiştiği bulunmuştur. Sonuç olarak araştırmamıza dahil olan kümeslerden elde edilen antikor titreleri ve % CV değerlerine bakıldığında; yüksek antikor titreleri-yüksek %CV değerleri veya düşük antikor titreleri-yüksek %CV değerleri, halen ülkemizde aşılama ile ilgili sorunların varlığını göstermektedir. Bu durum kümeslerde arzulanan homojen antikor titrelerine ulaşılmaması nedeniyle, kanatlı sürülerini enfeksiyon riskiyle karşı karşıya getirebileceği kanaatindeyiz.

Infectious Bronchitis Virus in Egypt: Genetic Diversity and Vaccination Strategies.

Infectious bronchitis virus (IBV) is a highly evolving avian pathogen that has increasingly imposed a negative impact on poultry industry worldwide. In the last 20 years, IBV has been continuously circulating among chicken flocks in Egypt causing huge economic losses to poultry production. Multiple IBV genotypes, namely, GI-1, GI-13, GI-16, and GI-23 have been reported in Egypt possessing different genetic and pathogenic features. Different vaccine programs are being used to control the spread of the disease in Egypt. However, the virus continues to spread and evolve where multiple IBV variants and several recombination evidence have been described. In this review, we highlight the current knowledge concerning IBV circulation, genesis, and vaccination strategies in Egypt. In addition, we analyze representative Egyptian IBV strains from an evolutionary perspective based on available data of their S1 gene. We also provide insight into the importance of surveillance programs and share our perspectives for better control of IBV circulating in Egypt.

Live Attenuated Infectious Bronchitis Virus Vaccines in Poultry: Modifying Local Viral Populations Dynamics.

Simple SummaryInfectious bronchitis (IB) is one of the more prevalent diseases in poultry, and it is caused by a virus belonging to the Coronaviridae family, the infectious bronchitis virus (IBV), a Gammacoronavirus which is related to the Betacoronavirus SARSCov-2 causing COVID-19 in humans. IB is mainly controlled by biosecurity and vaccines, although, it is a very challenging issue because the viral populations are constantly evolving by several factors. One of these factors is the same vaccines used for IB control, this could explain by recombination, reversion to virulence, or by favoring virus serotype selection. Thus, a human role in the change of viral populations can be identified by the IBV vaccine usage, this must be considered to achieve effective IB control.Infectious bronchitis virus (IBV) remains one of the most important diseases impacting poultry today. Its high adaptive capacity, attributable to the high mutation rate associated with its ssRNA(+), is one of its more important features. While biosecurity procedures and barriers have been shown to be preponderant factors in minimizing the impact of infectious bronchitis (IB), the environment and procedures associated with intensive poultry systems greatly influence the viral population dynamics. High-density poultry flocks facilitate recombination between different viruses, and even with live attenuated vaccines, which can change the dominant circulating field strains. Furthermore, the remaining issue of reversion to virulence gives rise to significant problems when vaccinal strains are introduced in places where their pathogenic variants have not been reported. Under specific conditions, live attenuated vaccines could also change the frequency of circulating viruses and enable replacement between different field strains. In summary, under a comprehensive approach, while vaccination is one of the most essential tools for controlling IB, the veterinarians, farmers, and official services role in its usage is central to minimizing alteration in a malleable viral population. Otherwise, vaccination is ultimately counterproductive.

Construction of an infectious bronchitis virus vaccine strain carrying chimeric S1 gene of a virulent isolate and its pathogenicity analysis.

Infectious bronchitis virus (IBV) is a member of genus gamma-coronavirus in the family Coronaviridae, causing serious economic losses to the poultry industry. Reverse genetics is a common technique to study the biological characteristics of viruses. So far, there is no BAC reverse genetic system available for rescue of IBV infectious clone. In the present study, a new strategy for the construction of IBV infectious cDNA clone was established. The full-length genomic cDNA of IBV vaccine strain H120 was constructed in pBAC vector from four IBV fragment subcloning vectors by homologous recombination, which contained the CMV promoter at the 5′ end and the hepatitis D virus ribozyme (HDVR) sequence and bovine growth hormone polyadenylation (BGH) sequence after the polyA tail at the 3′ end of the full-length cDNA. Subsequently, using the same technique, another plasmid pBAC-H120/SCS1 was also constructed, in which S1 gene from IBV H120 strain was replaced with that of a virulent SC021202 strain. Recombinant virus rH120 and rH120/SCS1 were rescued by transfecting the plasmids into BHK cells and passaged in embryonated chicken eggs. Finally, the pathogenicity of both the recombinant virus strains rH120 and rH120/SCS1 was evaluated in SPF chickens. The results showed that the chimeric rH120/SCS1 strain was not pathogenic compared with the wild-type IBV SC021202 strain and the chickens inoculated with rH120/SCS1 could resist challenge infection by IBV SC021202. Taken together, our results indicate that BAC reverse genetic system could be used to rescue IBV in vitro and IBV S1 protein alone might not be the key factor for IBV pathogenicity.Key points • BAC vector was used to construct IBV full-length cDNA by homologous recombination. • Based on four subcloning vectors, a recombinant chimeric IBV H120/SCS1 was constructed and rescued. • Pathogenicity of H120/SCS1 was similar to that of H120, but different to that of SC021202.

Genomic Stability for PCR Detection of Infectious Laryngotracheitis Virus and Infectious Bronchitis Virus in Poultry Dust Samples Stored Under Different Conditions.

Dust collected from the poultry house has been increasingly used as a population-level sample to monitor the presence of pathogens or to evaluate the administration of live vaccines. However, there are no guidelines for the storage of this sample type. This study investigated the stability of infectious laryngotracheitis virus (ILTV), a DNA virus, and infectious bronchitis virus (IBV), an RNA virus, in poultry dust kept under temperature and moisture conditions that mimic on-farm and laboratory storage. Dust samples were collected from chicks spray vaccinated with a live IBV vaccine and inoculated with a field ILTV strain via eye drop. Samples were stored under different moisture conditions (dry = 2% moisture, moist = 22%-71% moisture) and temperatures (-20, 4, 25, and 37 C) for different durations (0, 7, and 14 days, and 1, 2, 3, and 4 mo) in a factorial arrangement, followed by quantitative PCR for detection of virus genome copies (GC). The length of storage, moisture level, and storage temperature affected the viral genome load for ILTV and IBV but did not affect the number of positive samples for each virus. All treatment combinations were ILTV positive for at least 4 mo. In dry dust samples, all storage temperatures or durations had quantifiable ILTV or IBV GC. Moisture addition had a detrimental effect on viral genome load, causing an overall reduction of 0.3 log 10 for ILTV GC (7.29 and 6.97 log 10, P = 0.0001), and 1.3 log 10 for IBV GC (5.95 and 4.66 log 10, P = 0.0001), which are unlikely to have biologic significance. In conclusion, dry dust can be stored at any temperature up to 37 C for at least 4 mo without loss in qPCR detection of ILTV or IBV GC. Collection or storage of moist dust should be avoided, or air drying prior to storage is recommended if only moist dust is available.

Pathogenic characteristics of a QX-like infectious bronchitis virus strain SD in chickens exposed at different ages and protective efficacy of combining live homologous and heterologous vaccination

Continued reports of infections with infectious bronchitis virus (IBV) variants have occurred since its first isolation in the 1930s. Currently, QX-like IBVs are the predominant circulating genotype around the world. Here, the pathogenicity of QX-like IBV strain SD was characterized in chickens at different ages of exposure to the virus, and the protection efficacy of available vaccine combinations against IBV was evaluated. The results revealed that QX-like IBV strain SD was severely pathogenic in chickens, causing respiratory, urinary and reproductive infections, irrespective of age, based on clinical observations, viral distribution in tissues and a ciliostasis study. Severe respiratory signs, tracheal cilia injury, nephritis and abnormal development of the oviduct and ovarian follicles were evident throughout the experiment. A challenge experiment demonstrated that the homologous QX vaccine showed superior protection efficacy compared with other available vaccines, confirming the importance of IBV vaccine seed homology against the circulating IBV strains. Our findings aid an understanding of the pathogenicity of QX-like IBVs that may help to further control the infection.

Comparative Protective Efficacies of Novel Avian Paramyxovirus-Vectored Vaccines against Virulent Infectious Bronchitis Virus in Chickens.

Viral vectored vaccines are desirable alternatives for conventional infectious bronchitis virus (IBV) vaccines. We have recently shown that a recombinant Newcastle disease virus (rNDV) strain LaSota expressing the spike (S) protein of IBV strain Mass-41 (rLaSota/IBV-S) was a promising vaccine candidate for IBV. Here we evaluated a novel chimeric rNDV/avian paramyxovirus serotype 2 (rNDV/APMV-2) as a vaccine vector against IBV. The rNDV/APMV-2 vector was chosen because it is much safer than the rNDV strain LaSota vector, particularly for young chicks and chicken embryos. In order to determine the effectiveness of this vector, a recombinant rNDV/APMV-2 expressing the S protein of IBV strain Mass-41 (rNDV/APMV-2/IBV-S) was constructed. The protective efficacy of this vector vaccine was compared to that of the rNDV vector vaccine. In one study, groups of one-day-old specific-pathogenic-free (SPF) chickens were immunized with rLaSota/IBV-S and rNDV/APMV-2/IBV-S and challenged four weeks later with the homologous highly virulent IBV strain Mass-41. In another study, groups of broiler chickens were single (at day one or three weeks of age) or prime-boost (prime at day one and boost at three weeks of age) immunized with rLaSota/IBV-S and/or rNDV-APMV-2/IBV-S. At weeks six of age, chickens were challenged with a highly virulent IBV strain Mass-41. Our challenge study showed that novel rNDV/APMV-2/IBV-S provided similar protection as rLaSota/IBV-S in SPF chickens. However, compared to prime-boost immunization of chickens with chimeric rNDV/APMV-2, rLaSota/IBV-S and/or a live IBV vaccine, single immunization of chickens with rLaSota/IBV-S, or live IBV vaccine provided better protection against IBV. In conclusion, we have developed the novel rNDV/APMV-2 vector expressing S protein of IBV that can be a safer vaccine against IB in chickens. Our results also suggest a single immunization with a LaSota vectored IBV vaccine candidate provides better protection than prime-boost immunization regimens.

Limited Cross-Protection against Infectious Bronchitis Provided by Recombinant Infectious Bronchitis Viruses Expressing Heterologous Spike Glycoproteins

Gammacoronavirus infectious bronchitis virus (IBV) causes an economically important respiratory disease of poultry. Protective immunity is associated with the major structural protein, spike (S) glycoprotein, which induces neutralising antibodies and defines the serotype. Cross-protective immunity between serotypes is limited and can be difficult to predict. In this study, the ability of two recombinant IBV vaccine candidates, BeauR-M41(S) and BeauR-4/91(S), to induce cross-protection against a third serotype, QX, was assessed. Both rIBVs are genetically based on the Beaudette genome with only the S gene derived from either M41 or 4/91, two unrelated serotypes. The use of these rIBVs allowed for the assessment of the potential of M41 and 4/91 S glycoproteins to induce cross-protective immunity against a heterologous QX challenge. The impact of the order of vaccination was also assessed. Homologous primary and secondary vaccination with BeauR-M41(S) or BeauR-4/91(S) resulted in a significant reduction of infectious QX load in the trachea at four days post-challenge, whereas heterologous primary and secondary vaccination with BeauR-M41(S) and BeauR-4/91(S) reduced viral RNA load in the conjunctiva-associated lymphoid tissue (CALT). Both homologous and heterologous vaccination regimes reduced clinical signs and birds recovered more rapidly as compared with an unvaccinated/challenge control group. Despite both rIBV BeauR-M41(S) and BeauR-4/91(S) displaying limited replication in vivo, serum titres in these vaccinated groups were higher as compared with the unvaccinated/challenge control group. This suggests that vaccination with rIBV primed the birds for a boosted humoral response to heterologous QX challenge. Collectively, vaccination with the rIBV elicited limited protection against challenge, with failure to protect against tracheal ciliostasis, clinical manifestations, and viral replication. The use of a less attenuated recombinant vector that replicates throughout the respiratory tract could be required to elicit a stronger and prolonged protective immune response.

Characterization of novel, pathogenic field strains of infectious bronchitis virus (IBV) in poultry in Trinidad and Tobago.

Avian coronaviruses, including infectious bronchitis virus (IBV) and turkey coronavirus (TCoV), are economically important viruses affecting poultry worldwide. IBV is responsible for causing severe losses to the commercial poultry sector globally. The objectives of this study were to identify the viruses that were causing outbreaks of severe respiratory disease in chickens in Trinidad and Tobago (T&T) and to characterize the strains. Swab samples were collected from birds showing severe respiratory signs in five farms on the island of Trinidad. Samples were tested for the presence of IBV, as well as avian influenza virus (AIV), Newcastle disease virus (NDV) and avian metapneumovirus (aMPV) by real-time reverse transcription polymerase chain reaction (qRT-PCR). All samples from the five farms tested negative for AIV, NDV and aMPV; however, samples from clinically affected birds in all five of the farms tested positive for IBV. Genetic data revealed the presence of TCoV in chickens on two of the farms. Interestingly, these two farms had never reared turkeys. Phylogenetic analysis showed that IBV S1 sequences formed two distinct clusters. Two sequences grouped with vaccine strains within the GI-1 lineage, whereas three sequences grouped together, but separately from other defined lineages, forming a likely new lineage of IBV. Pairwise comparison revealed that the three unique variant strains within the distinct lineage of IBV were significantly different in their S1 nucleotide coding regions from viruses in the closest lineage (16% difference) and locally used vaccine strains (>20% difference). Results also suggested that one of the samples was a recombinant virus, generated from a recombination event between a Trinidad virus of the GI-1 lineage and a Trinidad virus of the newly defined lineage. Many amino acid differences were also observed between the S1 coding regions of the circulating field and vaccine strains, indicating that the IBV vaccines may not be protective. Vaccine-challenge studies are however needed to prove this.

The Construction and Immunoadjuvant Activities of the Oral Interleukin-17B Expressed by Lactobacillus plantarum NC8 Strain in the Infectious Bronchitis Virus Vaccination of Chickens.

Interleukin-17B (IL-17B) is a protective cytokine of the IL-17 family and plays an essential role in the regulation of mucosal inflammation. However, little is known about the role of IL-17B in the control of viral infections. In this study, a recombinant Lactobacillus plantarum, designated as NC8-ChIL17B, was constructed to express the chicken IL-17B (ChIL-17B) gene. The recombinant ChIL17B (rChIL17B) protein was about 14 kDa and was anchored to the surface of NC8 cells. In vitro, it was found that the rChIL17B protein inhibited the proliferation of the infectious bronchitis virus (IBV) through activation of nuclear factor kappa B (NF-κB) and the JAK (Janus kinase)-STAT (signal transducers and activators of transcription) signaling. Moreover, to evaluate the immunoadjuvant activities of NC8-ChIL17B, 40 three-day-old specific pathogen-free (SPF) chickens were divided into four groups. Three groups were orally vaccinated with fresh NC8, NC8-ChIL17B, and phosphate buffered saline (PBS), along with the infectious bronchitis virus vaccine, and the other group was the PBS-negative control. The results of the IBV-specific antibody titer and the concentration of the cytokines IL-2, IL-4, IL-6, and interferon gamma (IFN-γ) in sera, as well as the concentration of secretory immunoglobulin A (sIgA) in the tracheal and small intestinal mucosa, the number of cluster of differentiation 4 positive (CD4+) and cluster of differentiation 8 positive (CD8+) T cells in the blood, and the expression of immune-related genes all indicated that NC8-ChIL17B efficiently enhanced the humoral and cellular immune responses to IBV vaccine. Moreover, the viral loads in the NC8-ChIL17B- and IBV-vaccinated group were significantly lower than in the control groups, suggesting a significant promotion of the immunoprotection of IBV vaccination against the virulent IBV strain. Therefore, ChIL-17B is a promising, effective adjuvant candidate for chicken virus vaccines.