Evaluation of the effectiveness of two infectious bronchitis virus vaccine programs for preventing disease caused by a California IBV field isolate

Infectious bronchitis virus CA99 serotype was isolated from several broiler flocks in Northern California. The virus caused late-onset respiratory disease and increased airsacculitis condemnation in affected flocks despite the use of an established infectious bronchitis virus vaccination program. An experimental study compared Holland/Arkansas and Massachusetts/Arkansas vaccination protocols to determine the efficacy of commercial infectious bronchitis virus vaccines in reducing respiratory disease and airsacculitis lesions found at processing that were associated with a CA99 field isolate. All vaccination groups were given Massachusetts/Connecticut strains of infectious bronchitis virus vaccines at age 1 day followed by vaccination with either Holland/ Arkansas or Massachusetts/Arkansas vaccine strains at 18 days of age. Birds were challenged at age 31 days with a CA99 field isolate. Gross pathology, histopathology, and virus isolation were evaluated. Chickens vaccinated with Holland/Arkansas had marginally better protection against CA99 challenge than chickens vaccinated with Massachusetts/Arkansas, although differences were not statistically significant.     

Reciprocal Antibody and Complement Responses of Two Chicken Breeds to Vaccine Strains of Newcastle Disease Virus, Infectious Bursal Disease Virus and Infectious Bronchitis Virus

Serum antibody responses and haemolytic complement activity were evaluated in White Leghorn (WLH) and Rhode Island Red (RIR) chickens that were vaccinated with live-attenuated vaccines of Newcastle disease virus, or infectious bronchitis virus, or infectious bursal disease virus by means of ocular challenge at 10 times the normal vaccination dose. Complement titres in non-vaccinated birds were significantly higher in WLH birds compared to RIR birds. The lentogenic viral infection resulted in an immediate stimulation of complement activity, followed by a decrease to initial complement levels within 2 weeks post vaccination, when the antibody response took over immune defence. As compared to WLH chickens, RIR birds mounted a faster and significantly higher antibody response to the vaccine viruses used. In WLH hens, significantly higher haemolytic complement activity post vaccination was found as compared to RIR hens. Possible consequences of the observed differences in immune responsiveness of the two breeds to viral vaccines are discussed.     

The effects of avian leukosis virus subgroup J on broiler chicken performance and response to vaccination

The effects of avian leukosis virus subgroup J (ALV-J) infection on meat-type chickens reared in a simulated commercial setting were evaluated. Each of three ALV-J isolates was evaluated with both simulated horizontal transmission (SHT) and simulated vertical transmission (SVT). Mortality, morbidity, disease condemnations, and feed conversions were increased and body weights at processing were decreased in ALV-J infected birds as compared to sham inoculated hatch mates. The adverse effects of ALV-J infection were more severe in birds exposed by SVT than in birds exposed by SHT. At 8 weeks of age response to vaccination for infectious bronchitis virus and Newcastle disease virus or prior exposure to a pathogenic reovirus was assessed in the ALV-J and sham inoculated broiler chickens by challenge studies. Although not statistically significant, an overall trend of decreased protection to challenge after vaccination, or prior exposure, was observed in the ALV-J inoculates as compared to sham inoculated hatch mates. Differences in vaccine response were most evident in groups inoculated with ALV-J by the SVT route.     

Development and application of a multiplex polymerase chain reaction for avian respiratory agents

A multiplex polymerase chain reaction (PCR) was developed and optimized to simultaneously detect 6 avian respiratory pathogens. Six sets of specific oligonucleotide primers for infectious bronchitis virus (IBV), avian influenza virus (AIV), infectious laryngotracheitis virus (ILTV), Newcastle disease virus (NDV), Mycoplasma gallisepticum (MG), and Mycoplasma synoviae (MS) were used respectively in the test. With the use of agarose gel electrophoresis for detection of the PCR-amplified DNA products, the sensitivity of detection was between 10 pg for IBV, AIV, MG, and ILTV and 100 pg for NDV and MS after 35 cycles of PCR. Similar sensitivity of these primers was achieved with chickens experimentally infected with respiratory pathogens. In experimental infections, the multiplex PCR was able to detect all the infected chickens in each group at I and 2 wk postinfection as compared with serologic tests at 2 wk postinfection that confirmed the presence of specific antibodies. The multiplex PCR was also able to detect and differentiate coinfections with two or more pathogens. No specific DNA amplification for respiratory avian pathogens was observed among noninoculated birds kept separately as a negative control group.     

Infectious bronchitis virus nucleoprotein specific CTL response is generated prior to serum IgG

Infectious bronchitis (IB) is an acute and highly contagious viral respiratory disease of chickens. To understand the kinetics and relationships between the humoral (Ab) and antigen specific T cell immunity as well as pathological changes during infectious bronchitis virus (IBV) infection and immunization, one-week-old SPF chickens were vaccinated with live IBV H52 strain and challenged with IBV M41 15 days post primary infection. Chickens were sacrificed every 3 days to monitor antigen specific serum IgG and IBV nucleoprotein-specific immune responses using a chicken MHC I tetramer developed in our laboratory. The results demonstrated that T cell responses developed more rapidly than the humoral (Ab) immune response after vaccination with H52. However, serum IgG dramatically increased after M41 challenge. Chickens from the control, non-vaccinated group developed severe respiratory symptoms and demonstrated significant pathological changes in lung, kidney and bursa of Fabricius post challenge with M41. However, chickens vaccinated with H52 did not demonstrate clinical signs or histological changes post challenge with M41. These results indicated that the live IBV H52 inoculation effectively protected chickens from morbidity and pathological changes associated with IBV infection. These data facilitates the design of a new generation of IBV vaccine.     

Attenuation of avian infectious bronchitis virus by cold-adaptation.

Avian infectious bronchitis virus (IBV) Arkansas-type DPI strain was passaged 10 times in specific-pathogen-free (SPF) chicken embryos incubated at 28 C and 37 C. Virus grown at 28 C acquired cold-adapted (CA) and temperature-sensitive (TS) characteristics based on more-rapid growth at 28 C and a reduced ability to grown at 41 C, respectively, compared with non-cold-adapted (non-CA) virus grown at 37 C. The pathogenicity and immunogenicity were determined for CA and non-CA IBV in 1-day-old SPF chickens following intratracheal inoculation. The percentage of CA IBV-vaccinated chicks exhibiting respiratory disease exceeded 30% on only 1 day postinoculation (PI) (day 5 PI), compared with 8 days (days 2-9 PI) for birds given non-CA IBV. Mortality was 0% for CA IBV-vaccinated chickens and 6% for non-CA virus-vaccinated chickens. Microscopically, both CA and non-CA IBV caused diffuse tracheal deciliation, although mucosal hyperplasia, necrosis, and heterophil infiltration were more severe with non-CA IBV. Virus was reisolated from kidneys of chickens given CA IBV, suggesting the loss of the TS property. The instability of the TS property was confirmed by growth of the reisolated virus at 41 C. Both CA and non-CA viruses induced complete protection against homologous challenge virus infection of the upper respiratory tract.     

Effects of infectious bursal disease on Marek's disease vaccination: suppression of antiviral immune response

Studies were made to determine whether infectious bursal disease virus (IBDV) infection would affect the response of chickens to turkey herpesvirus (HVT) vaccination in the development and level of HVT viremia and virus-neutralizing (VN) antibodies to HVT. The HVT viremia in the vaccinated chickens was not affected by IBDV, whether IBDV was inoculated simultaneously with HVT vaccination at one day of age or whether it was inoculated 3 weeks postvaccination with HVT. However, VN antibody response to HVT was significantly suppressed (P less than 0.001) when vaccinated chickens were exposed to IBDV either at the time of vaccination or at 3 weeks postvaccination. Such immunosuppression by IBDV of VN antibody response to HVT vaccination may result in a reduced antiviral immunity against Marek's disease virus.     

Safety and efficacy of the cell-associated vaccine prepared by an attenuated infectious laryngotracheitis virus.

The safety and efficacy of the cell-associated (C-A) vaccine prepared by chicken embryo fibroblast (CEF) cells infected with the tissue-culture-modified strain of infectious laryngotracheitis (ILT) virus were studied in chickens. Over seventy percent of chickens inoculated with the C-A vaccine by the subcutaneous (S.C.) or intramuscular (I.M.) route at 1 day of age was protected against challenge with a virulent strain of ILT virus without any clinical signs. Chickens vaccinated with the C-A vaccine at 1 day of age acquired immunity within 6 days after vaccination, and the protection rate maintained more than 60% until 10 weeks post-vaccination. The C-A vaccine was invariably effective for chickens at various age. There was no evidence that the development of immunity was hindered by further vaccination with Newcastle disease and infectious bronchitis combined live vaccine. In addition, the C-A vaccine was safe when chickens were inoculated with 10 doses. In the field trials of the C-A vaccine, no adverse reaction was observed, and over 65% of vaccinated chickens was protected against the challenge of the virulent ILT virus at 8 weeks after vaccination.     

Effect of temperature-sensitive Mycoplasma gallisepticum vaccine preparations and routes of inoculation on resistance of white leghorns to challenge

One-week-old chickens were vaccinated with live or formalin-killed temperature-sensitive (TS) Mycoplasma gallisepticum (MG) either intranasally (IN) or subcutaneously (SQ). Live TS MG protected chickens against S6 strain challenge directly into the air sacs, regardless of route of vaccination. Killed MG, however, protected chickens only when administered SQ. Antibody to MG was detected in sera and in the tracheal and air-sac washings of only the chickens given live vaccine IN. The antibody present in tracheal and air-sac washings may be one of the mechanisms that play a role in resistance to MG challenge.     

Avian infectious bronchitis: cross-protection studies using different Australian subtypes

The cross-immunity of vaccinated chickens after challenge with some Australian infectious bronchitis viruses was assessed by humoral antibody responses and by ciliary activity in tracheal rings of vaccinated chickens following challenge. Four viruses were used for vaccination: Vac 3, Vac 4, both current infectious bronchitis vaccine viruses, and Q1/76 and N2/62. IBV N1/62 (synonym T0 and infectious bronchitis virus N9/74 (synonym Appin) were used to challenge the vaccinated chickens. Results showed a lack of correlation between humoral antibody levels and protection. Cross-immunity was found after vaccination with each subtype, but was lower for Vac 3 and Vac 4 than for Q1/76 and N2/62.     

Serological comparison and antigenic relationships of seven serotypes of infectious bronchitis virus using the hemagglutination-inhibition test

The antigenic relationships, antigenic spectrum, and immunogenicity of seven isolates of infectious bronchitis virus (IBV) were examined using the hemagglutination-inhibition (HI) test. Because there was a discontinuity of antigenic relationships and a high degree of cross-reactivity among serotypes of IBV in cross-hemagglutination-inhibition tests, the range of antigenic spectrum used to group the serotypes with the HI test should be wider than the limits suggested by the plaque-reduction test. The HI test may provide valuable information in monitoring the immune status of a flock following vaccination when the area has a history of infectious bronchitis infection. It may also be used as a rapid diagnostic test if a flock is experiencing an outbreak of a disease caused by emergence of a new type of IBV. Interpretation of HI titers in evaluating immune status of chickens following infection with IBV depends on further cross-challenge and cross-protection studies of various types of IBV.     

Protection of chickens after live and inactivated virus vaccination against challenge with nephropathogenic infectious bronchitis virus PA/Wolgemuth/98

Protection provided by live and inactivated virus vaccination against challenge with the virulent nephropathogenic infectious bronchitis virus (NIBV) strain PA/Wolgemuth/98 was assessed. Vaccinations with combinations of live attenuated strains Massachusetts (Mass) + Connecticut (Conn) or Mass + Arkansas (Ark) were given by eyedrop to 2-wk-old specific-pathogen-free leghorn chickens. After live infectious bronchitis virus (IBV) vaccination, some chickens at 6 wk of age received an injection of either an oil emulsion vaccine containing inactivated IBV strains Mass + Ark or an autogenous vaccine prepared from NIBV PA/Wolgemuth/98. Challenge with PA/Wolgemuth/98 was given via eyedrop at 10 wk of age. Serum IBV enzyme-linked immunosorbent assay antibody geometric mean titers (GMTs) after vaccination with the combinations of live attenuated strains were low, ranging from 184 to 1,354, prior to NIBV challenge at 10 wk of age. Both inactivated vaccines induced an anamnestic response of similar magnitudes with serum GMTs of 6,232-12,241. Assessment of protection following NIBV challenge was based on several criteria virus reisolation from trachea and kidney and renal microscopic pathology and IBV-specific antigen immunohistochemistry (IHC). Live attenuated virus vaccination alone with combinations of strains Mass + Conn or Mass + Ark did not protect the respiratory tract and kidney of chickens after PA/Wolgemuth/98 challenge. Chickens given a live combination vaccination of Mass + Conn and boosted with an inactivated Mass + Ark vaccine were also susceptible to NIBV challenge on the basis of virus isolation from trachea and kidney butshowed protection on the basis of renal microscopic pathology and IHC. Live IBV-primed chickens vaccinated with an autogenous inactivated PA/Wolgemuth/98 vaccine had the highest protection against homologous virulent NIBV challenge on the basis of virus isolation.     

Response of chickens to inoculation with a temperature-sensitive mutant of Mycoplasma gallisepticum

Newly hatched chickens were inoculated intranasally with either the S6 or TS 100 strain of Mycoplasma gallisepticum (MG) or they were left uninoculated. The three groups of chickens did not differ discernibly in body, spleen, or bursa weight during the 27-day sampling period. However, the S6-inoculated chickens showed a more pronounced cellular response in the nasal passages and had nearly complete lymphoid depletion in the spleens. The TS 100-inoculated birds expressed only a mild cellular reaction, which was localized in the nasal passages. Uninoculated chickens appeared normal histologically. Serologic tests such as rapid serum plate agglutination, hemagglutination-inhibition, and radioimmunoassay were able to detect antibody responses of chickens to MG inoculations yet could not differentiate the response to TS 100 from the response to S6. Tracheal secretions in intact TS 100-inoculated chickens contained antibodies to MG, yet only one-half of the bursectomized inoculated chickens contained detectable antibody, which appeared to be IgG. This led to the conclusion that bursectomy suppresses the appearance of locally synthesized IgG antibodies to MG in tracheal washings. The locally produced antibody was considered important in the development of resistance induced by intranasal inoculation of TS mutants.     

Humoral immune responses to chicken infectious anemia virus in three strains of chickens in a closed flock

This is a comparative study on seroconversion to chicken infectious anemia virus (CIAV) in a closed flock of specific-pathogen-free chickens undergoing a natural outbreak and after vaccination of some of these flocks with a commercial, live vaccine. The N2a strain (B21B21 haplotype) had the highest seroconversion after natural infection (94%) or vaccination (100%), followed by the P2a strain (B19B19) at 75%-82% seroconversion after natural infection and 85% seroconversion after vaccination. The S13 (B13B13) chickens were 26% seropositive after natural infection and 75% seropositive after vaccination. N2a chickens with polymerase chain reaction (PCR)-positive tissues were 97% seropositive compared to 80%-83% PCR-positive and seropositive for the P2a chickens and only 8% seropositive and PCR-positive for the S13 chickens. Seroconversion occurred at or near sexual maturity after natural infection in seven flocks studied.     

The resistance to reinfection of tracheal organ cultures from chickens previously infected with avian infections bronchitis virus.

Lower titres of avian infectious bronchitis (AIB) virus were found following the infection of tracheal organ cultures prepared from chickens that had been given AIB virus intranasally six weeks previously than were found following the infection of organ cultures prepared from untreated or from passively-immune chickens. No infectious virus was found in the tracheal organ cultures at the time they were prepared.     

Immune responses of breeding chickens to trivalent oil emulsion vaccines: responses to infectious bronchitis

Three similar flocks of broiler breeder parent chickens that had been given live infections bronchitis (IB) vaccines during rearing were injected at 20 weeks of age with three different oil emulsion vaccines: a commercial monovalent Newcastle disease (ND) vaccine (flock A); an experimental bivalent vaccine containing ND and infectious bursal disease (IBD) components (flock B); and an experimental trivalent vaccine containing ND, IBD and IB components (flock C). One week after vaccination 40 hens from flock A and 40 from flock C were taken to the laboratory and their egg yields individually recorded. At 37 weeks of age they were challenged by aerosol exposure to virulent IB virus. The egg production dropped significantly in the hens from flock A but not in the hens from flock C. On the farm, flock C showed a higher mean IB virus antibody titre four weeks after vaccination but titres rose in all three flocks indicating the presence of active IB virus infection. No differences in egg yields were found between the three farm flocks.     

CAV与REV共感染SPF鸡对疫苗免疫反应的抑制作用

用1日龄SPF鸡人工感染鸡贫血病毒(CAV)和禽网状内皮增生病病毒(REV),探讨病毒感染对鸡体疫苗免疫反应的影响。结果表明,在用禽流感病毒(AIV,H5和H9)疫苗免疫后,CAV与REV单独感染均显著抑制了鸡体对H5和H9亚型禽流感病毒灭活疫苗的HI抗体反应,在CAV与REV共感染后,这种抑制作用更为明显。CAV单独感染后鸡体对新城疫病毒(NDV)和传染性法氏囊病病毒(IBDV)疫苗的免疫反应受到抑制,但与对照组在统计学上的差异不显著,然而,CAV可以显著加重REV感染对鸡体在NDV和IBDV疫苗免疫后抗体反应的抑制作用。从而证实CAV与REV共感染在疫苗免疫抑制上有协同作用。     

Enzyme-linked immunosorbent assay for the detection of antibodies to infectious bronchitis virus

An enzyme-linked immunosorbent assay (ELISA) for antibodies to avian infectious bronchitis (IB) virus is described. The immune response of chickens following vaccination with IB virus was monitored using this test, and the titers were compared with those obtained by serum neutralization. The ELISA appears to be suitable for IB serology.     

Standardization and application of the enzyme-linked immunosorbent assay for infectious bronchitis

The indirect enzyme-linked immunosorbent assay (ELISA) was used to detect antibody to infectious bronchitis virus in chickens. The serum-neutralization test for infectious bronchitis (SNIB) was used as a reference serologic test. The ELISA proved to be useful for monitoring antibody responses following vaccination of leghorn chicks. The titers obtained with the ELISA and SNIB showed an overall correlation, but results suggest the involvement of different antibody classes.     

Ciliary activity: a criterion for associating resistance to infectious bronchitis virus infection with ELISA antibody titer

Vaccination and challenge experiments using infectious bronchitis virus (IBV) were conducted on groups of specific-pathogen-free chickens. Three weeks post-vaccination with one of the four IBV strains used, chickens were challenged with the homologous immunizing strain of IBV. Subsequently, the chickens were sacrificed, their tracheas were examined for ciliostasis, and the specific IBV antibody content of their sera was measured by enzyme-linked immunosorbent assay (ELISA). Results showed that protection was conferred by primary vaccination, as ciliostasis was not observed in tracheas from groups vaccinated and then challenged. No protection was observed in control groups that received only a challenge exposure, and the virus was readily recovered from their tracheas. Homologous protection was present in chickens that had ELISA titers as low as 624 and neutralization indices as low as 2.9, whereas susceptible controls had titers of less than 100 and less than 1.0, respectively.