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PubMed1999

Induction of protective immunity in chickens immunised with plasmid DNA encoding infectious bursal disease virus antigens.

Direct DNA inoculations were used to determine the efficacy of gene immunisation of chickens to elicit protective immune responses against infectious bursal disease virus (IBDV). The vp2 gene of IBDV strains GP40 and D78, and the vp2-vp4-vp3 encoding segment of strain D78 were cloned in an expression vector which consisted of human cytomegalovirus (HCMV) immediate early enhancer and promoter, adenovirus tripartite leader sequences and SV40 polyadenylation signal. For purification of vaccine-quality plasmid DNA from E. coli, an effective method was developed. Chickens were vaccinated by inoculation of DNA by two routes (intramuscular and intraperitoneal). Two weeks later, chickens were boosted with DNA, and at 2 weeks post-boost, they were challenged with virulent IBDV strain. Low to undetectable levels of IBDV-specific antibodies and no protection were observed with DNA encoding VP2. However, plasmids encoding VP2-VP4-VP3 induced IBDV-specific antibodies and protection in the chickens. DNA immunisation opens a new approach to the development of gene vaccines for chickens against infectious diseases.

Animals↗

PubMedDate not supplied

Molecular characterization of Spanish infectious bursal disease virus field isolates.

Nine Spanish isolates of infectious bursal disease virus (IBDV) were characterized and classified after reverse transcriptase-polymerase chain reaction of a 248-bp fragment of the VP2 gene hypervariable region and restriction fragment length polymorphism (RFLP). The restriction endonucleases (REs) used were BstNI, Sad, SspI, TaqI, DraI, and StyI. Sequencing of the amplified product and further comparison of these sequences with published sequence data from other IBDV strains were also performed. Very virulent and classic strains were identified. None of the strains identified had molecular characteristics similar to that of the American variant strains. Four very virulent strains (VG-248, 5939, 6145, and 7333) were digested by the TaqI, SspI, and StyI enzymes. The sequences of these strains were closely related to other European and Japanese very virulent IBDV (vvIBDV) strains. Strains VG-311, VG-262, and VG-208 were digested by the BstNI and Sad REs and were classified as classic strains. Strains VG-276 and VG-313 had unique RFLP patterns. VG-276 exhibited the SspI RE site, which has been reported as a characteristic of vvIBDV strains, whereas the VG-313 strain exhibited a Sad and StyI RE site indicative of the classic IBDV Edgar and 52-70 strains. However, nucleotide sequence analysis of the amplified hypervariable region strain VG-276 revealed a higher identity with the classic strains STC, 52/70, and 9109 IBDV strains, whereas strain VG-313 exhibited a higher identity with the vvIBDV strains.

Animals↗

PubMedDate not supplied

Virulent vaccine strains of infectious bursal disease virus not distinguishable from wild-type viruses with marker the use of a molecular.

Molecular techniques have not only made timely and accurate detection of infectious bursal disease viruses (IBDVs) possible but also have allowed the identification of viral strains. Previously, we identified a genetic marker that distinguished wild-type IBDV strains from vaccine strains of the virus. The marker was an NgoM IV restriction enzyme site in the VP2 gene that was present in 10 wild-type viruses but not 16 vaccine strains of IBDV. On the basis of that study, we concluded that the NgoM IV marker could be useful in the identification of wild-type potentially pathogenic strains of this virus. Because virulent (hot) vaccine strains of IBDV are used to vaccinate commercial poultry, it was important to determine if the NgoM IV marker was present in these virulent vaccines. The infectious bursal disease Blen and Bursa Vac virulent vaccines were examined and determined to contain the marker. We concluded that the presence of this marker was not unique to wild-type strains of the virus. The absence of the NgoM IV marker, however, was consistent with some level of attenuation, and its presence appears to be consistent with virulent IBDV strains.

Genetic Markers↗

PubMed2004

Heteroduplex mobility assay for genotyping infectious bursal disease virus.

A heteroduplex mobility assay (HMA) was developed to genotype infectious bursal disease virus (IBDV). This method analyzed 390-base pair (bp) polymerase chain reaction (PCR) products, encompassing the hypervariable region of the VP2 gene. IBDV strains from the United States and other countries were analyzed. The HMA was able to differentiate standard, antigenic variants and very virulent strains of IBDV. Minor differences between different strains from the same subtype were also detected. Close relationships between field IBDV with vaccines prepared with Delaware E strain were determined by HMA. The results obtained by HMA were confirmed by restriction fragment length polymorphism (RFLP) and phylogenetic analysis of nucleotide sequences. The HMA proved to be a useful technique to rapidly genotype different field strains of IBDV and should prove to be a useful tool in epidemiologic studies.

Genotype↗

PubMed2005

Development of a one-step strip test for the diagnosis of chicken infectious bursal disease.

A rapid diagnostic strip for chicken infectious bursal disease (IBD) was developed based on membrane chromatography using high-affinity monoclonal antibodies directed to chicken infectious bursal disease virus (IBDV). The diagnostic strip has high specificity for detection of chicken IBDV antigen and recognizes a variety of the virus isolates, including virulent and attenuated strains, with no cross-reactivity to other viruses, such as Newcastle disease virus, Marek's disease virus, infectious bronchitis virus, infectious laryngotracheitis virus, and egg-drop-syndrome virus. The results showed that its specificity was highly consistent with the agar-gel precipitation test (AGP). The diagnostic strip detected as low as 800 median egg lethal dose (ELD50) viruses in the IBDV BC6/85-infected sample, which was comparable with AC-ELISA (400 ELD50) and 32 times more sensitive than the AGP test (2.56 x 10(4) ELD50). In experimental infection, IBDV was detected in the bursa as early as 36 hr postinfection with the diagnostic strip before the clinical signs and gross lesions appeared. It takes only 1-2 min to do a strip test to detect chicken IBDV antigen after the specimen is grounded in a whirl pack with finger massage.

Animals↗

PubMed2005

Detection of infectious bursal disease virus in various lymphoid tissues of experimentally infected specific pathogen free chickens by different reverse transcription polymerase chain reaction assays.

Infectious bursal disease (IBD) is a worldwide distributed immunosuppressive viral disease in young chickens, controlled by vaccination. Emergence of several strains of IBD virus (IBDV) has created a demand for strain-specific diagnostic tools. In the present experiment, five different reverse transcription polymerase chain reaction (RT-PCR) assays, including two recently developed strain-specific assays, were employed for detection of ribonucleic acid (RNA) from three different IBDV strains in bursa tissue samples from experimentally infected specific pathogen free chickens. The virus strains included vaccine strain D78, classical strain Faragher 52/70, and the very virulent Danish strain DK01. The presence of the virus infection was confirmed by histopathologic evaluation of bursa lesions. The largest number of positive samples was obtained with a strain-specific two-step multiplex (MPX) RT-PCR assay based on iScript enzyme, and the commercially available Qiagen one-step RT-PCR. Between these methods, agreement was obtained for 57 of 59 samples. Because the Qiagen one-step RT-PCR assay was suggested as the more sensitive of these two assays, it was used for detection of IBDV in bone marrow, spleen, thymus, and cecal tonsils from experimentally infected chickens. The identity of the virus strains involved was confirmed by MPX RT-PCR. In conclusion, the MPX RT-PCR represented a reliable assay for detection and differentiation of IBDV strains in selected lymphoid tissues of chickens. All three of the IBDV strains used were detected in bursa tissues, whereas only the two virulent strains were detected in bone marrow, spleen, and thymus.

Animals↗

PubMed2006

Infectious bursal disease virus-induced immunosuppression exacerbates Campylobacter jejuni colonization and shedding in chickens.

Campylobacter jejuni is the leading cause of food-borne bacterial gastroenteritis in humans in the United States. Infectious bursal disease virus (IBDV) causes an immunosuppressive disease in young chickens. To analyze a possible role of IBDV-induced immunosuppression in colonization and shedding of C. jejuni, two experiments were conducted. In both experiments, group 1 consisted of noninoculated control chickens, groups 2 and 3 were inoculated with varying doses of C. jejuni, and groups 4 and 5 were inoculated initially with IBDV followed by doses of C. jejuni similar to groups 2 and 3. Campylobacter jejuni was recovered from the cloaca and cecum, but not the small intestines, from all chickens in groups 2 and 3. In groups 4 and 5, C. jejuni was recovered from the small intestines, cecum, and cloaca from all chickens. The amount (colony-forming units/sample) of C. jejuni recovered from chickens in groups 4 and 5 was significantly greater (P < 0.05) than the amount recovered from chickens in groups 2 and 3; and C. jejuni was also present sooner in these groups than in groups 2 and 3. Bursa samples from chickens in groups 4 and 5 were significantly smaller (P < 0.05) than in the other groups. Additionally, real-time polymerase chain reaction results for IBDV were positive in groups 4 and 5 and negative in all other groups. This study indicated that IBDV infection exacerbated colonization and shedding of C. jejuni, presumably through the immune suppression this virus causes in chickens. It highlights the need for further investigation into the role of immunosuppression in preharvest control strategies for food-borne disease-causing agents.

Animals↗

PubMed1977

Effect of infectious bursal disease on the severity of Eimeria tenella infections in broiler chicks.

A study was initiated to determine whether prior exposure to infectious bursal disease virus (IBDV) influenced the susceptibility of young broiler chicks to Eimeria tentella infections. When one-day-old chicks infected with IBDV were subsequently challenged with E. tenella at 7 days of age, they suffered significantly higher mortality and lesion scores than their hatchmates which were not exposed to IBDV. Initial exposure to IBDV also had an effect on the development of E. tenella induced hemorrhaging. Hemorrhaging commenced late on the third day following E. tenella inoculation in chicks infected with both IBDV and E. tenella, while bleeding did not occur in the E. tenella alone group until late on the fourth day post-coccidial challenge. No significant differences in weight gains at 7 or 14 days after E. tenella infection were observed between chicks receiving the double infection and those given only E. tenella.

Animals↗

PubMed1978

Vaccination against infectious bronchitis and the immunosuppressive effects of infectious bursal disease.

An immunosuppressive effect was demonstrated in chickens which were infected with infectious bursal disease virus (IBDV) early in life and prior to or shortly after vaccination with infectious bronchitis virus (IBV). This effect was evident by an increased susceptibility to respiratory tract infection with IBV and reduced virus-serum neutralizing antibody levels. Chickens which were hatched from dams susceptible to infectious bursal disease (IBD) were less responsive to IBV immunization attempts, if exposed to IBDV, than were those individuals hatched from IBD immune dams. However, in some cases, chickens from IBD immune dams were also more susceptible to IBV challenge when they had been exposed to IBDV and when compared to birds unexposed to IBDV but vaccinated against IB. An effect of cyclophosphamide on the bursa of Fabricius also had an immunosuppressive action on IBV immunity which was similar to the results from IBDV exposure. The data engendered from these trials may explain the unsatisfactory immunity sometimes observed under field conditions when broilers and replacement pullets are vaccinated at an early age.

Animals↗

PubMed1983

Vaccination against Marek's disease and infectious bursal disease. I. Development of a bivalent live vaccine by co-cultivating turkey herpesvirus and infectious bursal disease vaccine viruses in chicken embryo fibroblast monolayers.

Vaccine viruses of Marek's disease (MD), the FC-126 strain of the herpesvirus of turkeys (HVT), and infectious bursal disease (IBD), the Bursa-Vac-M strain of IBD virus (IBDV), were propagated in the same chicken embryo fibroblast (CEF) monolayers by superinfection. Co-infection of the two viruses in the same CEF culture or in a single cell can be demonstrated by staining with acridine orange and by electron microscopy.l This study was conducted with a superinfected live bivalent vaccine (HVT/IBDV) in one-day-old specific pathogen-free (SPF) and conventional White Leghorn (CWL) chicks. Chickens vaccinated with HVT/IBDV produced persistent HVT viremia for at least 6 weeks postvaccination; significantly higher IBD virus neutralizing antibody levels were observed for the entire 8-week experimental period when compared to unvaccinated controls. Differences between virus neutralizing antibody titers stimulated by HVT/IBDV and IBDV alone were not statistically significant in SPF chickens; however, due to maternal antibody there were some significant differences in CWL chickens. Both the HVT/IBDV bivalent vaccine and the monovalent IBDV vaccine protected the chickens against challenge with virulent IBDV. Based on microscopic lesions and bursa: body weight indexes, challenge of vaccinated chickens with virulent IBDV caused no atrophy in the bursa of Fabricius, but severe tissue destruction was observed in unvaccinated challenged controls. Chickens vaccinated with HVT/IBDV vaccine or with HVT alone were effectively protected against MD when compared to the controls.

Animals↗

PubMed2003

[Rapid construction of infectious clones of infectious bursal disease virus].

A rapid procedure was established for rescuing infectious bursal disease virus (IBDV), an important pathogen in poultry. A full-length cDNA clone of the segment B of a CEF-adapted IBDV strain HZ2 was constructed by long RT-PCR, and the 2827 bp nucleotide sequence, including the 5 - and 3 -noncoding regions (NCR), was established. Then the cDNA clone of segment B was engineered to make it contain three silent nucleotide changes, creating a new EcoRV site that was different from the parent virus sequences, by site-directed silent mutagenesis. Cotransfection of eukaryotic expression recombinants containing modified segment A and segment B with Lipofectamine into Vero cells resulted in the expression of IBDV RNA and proteins, as confirmed by Northern RNA dot hybridization and indirect immunofluorescence assay analysis. The change of cell morphology after cotransfection and passages of cell cultures was similar to that of cells infected by authentic IBDV, causing cellular pathogenic effects (CPE). The virus-like particles at 55-60 nm were observed under electron microscopy, affirming the rescue of IBDV. The genetic markers were retained in the recovered progeny virus.

Animals↗

PubMed1976

Immunodeficiency in the chicken. IV. An immunological study of infectious bursal disease.

Chickens inoculated orally with infectious bursal disease virus (IBDV) 1 day after hatching subsequently showed a 50% incidence of immunodeficiency but little mortality. Antibody responses against IBDV and to immunization with sheep red blood cells (SRBC) or human serum albumin (HSA) were suppressed. Serum IgG concentration was decreased while IgM occurred exclusively in its 7S monomeric form (mIgM). An allotypic marker of chicken IgM (Mla) was lacking in mIgM derived from IBDV-infected birds. The loss of Mla occurred gradually in several birds between 3 and 12 weeks after perinatal infection. Inoculation of IBDV into chickens 3 weeks after hatching resulted in 50% mortality level but little immunodeficiency. Paradoxically, the serum IgG concentration was elevated, in comparison with normal birds. Histology of the bursa showed permanent hypo- or aplasia of follicles irrespective of the age of infection. The results suggest that bursal but not peripheral B cells are targets for IBDV, and immunodeficiency results from impaired peripheral seeding of B cells in infected juvenile chickens.

Age Factors↗

PubMed1977

Effect of infectious bursal disease on the response of chickens to Mycoplasma synoviae, Newcastle disease virus, and infectious bronchitis virus.

At 35 days of age, chickens which as 1-day-old chicks were inoculated with the infectious bursal disease virus (IBDV) had significantly lower antibody titers against Mycoplasma synoviae, Newcastle disease virus, and infectious bronchitis virus than did those never inoculated with IBDV. The IBDV also had a marked effect on the development of air-sac lesions. Birds infected with IBDV that were later inoculated with M synoviae (day 14), Newcastle disease virus (days 14 and 28) experienced an increased incidence and greater seversity of airsacculitis than did chicks which were not exposed to IBDV.

Air Sacs↗

PubMedDate not supplied

Growth of infectious bursal disease virus with plaque formation in chick embryo fibroblast cell culture.

The WA69 isolant of infectious bursal disease virus (IBDV) induced cytopathic effects and plaque formation in chick embryo fibroblast (CEF) cultures after serial passages in embryonated eggs and then in CEF cultures. The plaque-forming agent was cloned (designated WA69 clone) and identified as IBDV on the basis of serologic response in inoculated birds and its antigenic relationship to other known IBDV isolants. The WA69 clone replicated rapidly in CEF cultures, reaching peak titers at 48 hours postinoculation, and the virus caused only minimal histologic lesions of the bursa when inoculated into 3-week-old chicks from a specific-pathogen-free flock. The growth of IBDV in CEF cultures with plaque formation may provide a simple in vitro system for virological and serological studies of IBDV.

Animals↗

PubMedDate not supplied

An immunoperoxidase monoclonal antibody stain for rapid diagnosis of infectious bursal disease.

Cell smears of chicken-embryo-fibroblast (CEF) cultures and bursa of Fabricius from chickens experimentally infected with six different strains of infectious bursal disease virus (IBDV) were examined for the presence of IBDV by the avidin-biotin-peroxidase complex method of immunoperoxidase (IP) staining using a monoclonal antibody specific for IBDV designated BK70. IBDV of different strains and serotypes were readily detected by the IP method in cell smears prepared from infected CEF cultures and from bursas. Bursal cells were positive for IP stain in most of the infected bursas (87.5%), despite their mild IBD lesions. Positive IP staining of bursal smears was well correlated with the recovery of IBDV from the bursas and with IBD lesions in the bursas. IP stain with a monoclonal antibody (BK70) appeared potentially useful for rapid and definitive diagnosis of IBD.

Animals↗

PubMedDate not supplied

Group and strain-specific neutralization sites of infectious bursal disease virus defined with monoclonal antibodies.

Two somatic cell hybridizations were performed utilizing splenocytes from mice immunized with one or more strains of infectious bursal disease virus (IBDV). Supernatants from hybridoma cell lines were initially screened by the enzyme-linked immunosorbent assay (ELISA) against multiple strains of IBDV. Cell lines that secreted antibodies with ELISA reactivity patterns of interest were cloned, and their monoclonal antibodies (MCAs) were subsequently tested in cross-virus-neutralization tests. Two of the nine MCAs selected exhibited strong neutralizing activity and precipitated IBDV antigens in agar gel precipitin tests as well. MCA B69 significantly neutralized only the cloned D78 strain of IBDV, whereas MCA R63 neutralized all IBDV strains (representing both serotype I and II viruses) against which it was tested. Results of competitive ELISAs that used the R63 and B69 MCAs showed that the two neutralization sites on the D78 strain were not overlapping.

Animals↗

PubMedDate not supplied

Comparison of a kinetic-based enzyme-linked immunosorbent assay (KELISA) and virus-neutralization test for infectious bursal disease virus. II. Decay of maternal antibody in progeny from white Leghorns receiving various vaccination regimens.

A computer-assisted single-serum-dilution indirect kinetic-based enzyme-linked immunosorbent assay (KELISA) was used for quantitating the natural decay rate of infectious bursal disease virus (IBDV) maternal antibody in progeny obtained from white leghorn breeders. The KELISA results were compared with those of a standard virus-neutralization (VN) test. Pullets were subjected to two IBDV immunization regimens. Group 1 was vaccinated at weeks 0, 2, and 10 with two live vaccines in drinking water and at week 20 with an oil-emulsified (SC) IBDV vaccine, group 2 received only the first and last immunizations, and group 3 served as the unvaccinated control. Pullets were artificially inseminated at 28 weeks of age. Progeny chicks from each group were bled every other day for 47 days. Both KELISA and VN test detected linear relationship in the decay of maternal antibodies. The VN test detected no significant differences (P greater than 0.05) in the IBDV maternal antibody titers at day 1 or in the rate of decay between the progeny from groups 1 and 2. The VN maternal antibody titers decreased at a rate of 0.16 log2 titer per day. In contrast, KELISA revealed higher IBDV maternal antibody titers in day-old progeny from pullets vaccinated 4 times (log2 = 14.3). However, KELISA titers of progeny from this group decreased at a faster rate than titers of progeny from pullets vaccinated twice (0.20 vs. 0.13 log2 titer per day).

Animals↗

PubMedDate not supplied

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.

Animals↗