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PubMed1982

Humoral and cell-mediated immune responses in chickens with infectious bursal disease.

Primary and secondary immune responses to Newcastle disease virus (NDV) was evaluated in chickens infected with infectious bursal disease virus (IBDV) at one and 28 days of age. The geometric mean primary hemagglutination-inhibition antibody titers (GMT) of chickens infected with IBDV at one day of age was significantly lower (P less than or equal to 0.01) than those infected at 28 days of age. Infection with IBDV had no influence on secondary immune response to NDV. The effect of IBDV infection at one day of age on the cell-mediated immunity of chickens was evaluated by skin allograft acceptance or survival time. There was no significant difference between the percentage of grafts accepted in IBDV infected and noninfected control chickens. However, the mean graft survival time in the IBDV infected chickens was significantly longer (P less than or equal to 0.05) than those in the control group. This suggested a suppression of cell-mediated immunity due to IBDV infection.

Animals↗

PubMed1998

A comparison of two RNA isolation methods for double-stranded RNA of infectious bursal disease virus.

Infectious bursal disease virus (IBDV), a member of the birnaviridae family, contains a bisegmented double-stranded RNA (dsRNA) genome. The segments are linked covalently at 5' termini by a large (90 kDa) viral genomic protein that migrates similar to viral RNA dependent RNA polymerase of IBDV. A proteinase K digestion based approach and acid-guanidium-phenol-chloroform (AGPC) RNA extraction method were used to extract dsRNA of IBDV from infected bursae. After extraction, dsRNA of IBDV was purified by precipitation with lithium chloride. The yield and purity of dsRNA of IBDV extracted by AGPC method was lower than that of proteinase K digestion based approach. This observation correlates with the presence of a genome-linked protein in IBDV. Although dsRNA obtained by both methods are suitable for reverse transcription-polymerase chain reaction (RT-PCR) amplification of at least up to 1201 base pairs (bp) of cDNA, dsRNA extracted by the proteinase K digestion method is more suitable than that by AGPC method for the amplification of longer fragments (1958 bp) of IBDV cDNA by PCR.

Animals↗

PubMed1999

Amplification and cloning of infectious bursal disease virus genomic RNA segments by long and accurate PCR.

Improved methods of reverse transcription, polymerase chain reaction (PCR) amplification, and cloning of full-length coding region of both strands of infectious bursal disease virus (IBDV) variant strain E genome were developed. Denaturation of IBDV RNA by heat in the presence of primers and use of a reverse transcriptase lacking RNase-H activity produced full-length coding region and partial non-coding region cDNA copies of the viral genomic segments. Digestion of the RNA component of RNA-cDNA hybrids by RNase-H followed by long and accurate PCR (LA-PCR) amplification of IBDV cDNA in a single step resulted in the synthesis of 3182 base-pairs (bp) of segment A and 2777 bp of segment B cDNA copies of IBDV genome. The resulting amplicons were successfully cloned and sequenced revealing their identity of IBDV. The LA-PCR method can be utilized for the amplification and cloning of the other IBDV strains or isolates and will greatly enhance the availability of sequence information or infectious cDNA copies of IBDV.

Animals↗

PubMed2000

Effects of hydrostatic pressure on the structure and biological activity of infectious bursal disease virus.

The effects of high hydrostatic pressure on the structure and biological activity of infectious bursal disease virus (IBDV), a commercially important pathogen of chickens, were investigated. IBDV was completely dissociated into subunits at a pressure of 240 MPa and 0 degrees C revealed by the change in intrinsic fluorescence spectrum and light scattering. The dissociation of IBDV showed abnormal concentration dependence as observed for some other viruses. Electron microscopy study showed that morphology of IBDV had an obvious change after pressure treatment at 0 degrees C. It was found that elevating pressure destroyed the infectivity of IBDV, and a completely pressure-inactivated IBDV could be obtained under proper conditions. The pressure-inactivated IBDV retained the original immunogenic properties and could elicit high titers of virus neutralizing antibodies. These results indicate that hydrostatic pressure provides a potential physical means to prepare antiviral vaccine.

Anilino Naphthalenesulfonates↗

PubMed1999

Evaluation of serological, histological and immunocytochemical methods for the detection of infectious bursal disease virus infection in broiler flocks in New Zealand.

AIMS: TO study and compare three diagnostic methods for the detection of infectious bursal disease virus (IBDV) infection. METHODS: Samples of sera and bursae were collected from two flocks from each of two broiler farms (Farms A and B) in which IBD had occurred or was suspected to have occurred. Sera were tested in ELISA and agar gel precipitation tests for the presence of IBD antibodies. Bursae were examined histologically for evidence of IBD lesions. An immunocytochemical test was developed to detect IBDV antigens in sections of bursa. RESULTS: Bursae from serologically negative, 45-day-old birds from Farm A, Flock 1 and from serologically positive 49-day-old birds from Farm B, Flock 1 had histological and immunocytochemical evidence of IBDV infection. Birds from Farm A, Flock 2, sampled 12 months after the sampling of Flock 1, and specific-pathogen-free birds, showed no evidence of IBDV infection by any of the three diagnostic methods. Birds from Farm B, Flock 2, sampled on four occasions, were positive for IBD at 20 days of age by histology and immunocytochemistry, but did not seroconvert until 42 days of age. CONCLUSIONS: Serological testing is not a reliable method for the detection of IBDV infection in New Zealand broiler flocks because antibodies may not have developed to detectable levels by the time of slaughter. Histological examination of bursae allowed the demonstration of IBD-like lesions, but these need to be differentiated from those caused by other agents. The immunocytochemistry test was able to detect early IBDV infection. It provided a rapid, definitive diagnosis and may be useful in control programmes. The results from Farm A demonstrate that strict biosecurity measures can be successful in the eradication of IBDV.

Journal Article↗

PubMed2001

Apoptosis is induced by infectious bursal disease virus replication in productively infected cells as well as in antigen-negative cells in their vicinity.

The kinetics of infectious bursal disease virus (IBDV) replication and induction of apoptosis were investigated in vitro and in vivo. After infection of chicken embryo (CE) cells with IBDV strain Cu-1, the proportion of apoptotic cells increased from 5.8% at 4 h post-infection (p.i.) to 64.5% at 48 h p.i. The proportion of apoptotic cells correlated with IBDV replication. UV-inactivated IBDV particles did not induce apoptosis. Double labelling revealed that, early after infection, the majority of antigen-expressing cells were not apoptotic; double-labelled cells appeared more frequently at later times. Remarkably, apoptotic cells were frequently located in the vicinity of antigen-expressing cells. This indicated that an apoptosis-inducing factor(s) might be released by cells that replicate IBDV. Since interferon (IFN) production has been demonstrated after IBDV infection, IFN was considered to be one of several factors. However, supernatants of infected CE cells in which virus infectivity had been neutralized were not sufficient to induce apoptosis. Similar results were observed in the infected bursae of Fabricius: early after infection, most of the cells either showed virus antigens or were apoptotic. Again, double-labelled cells appeared more frequently late after infection. This suggests that indirect mechanisms might also be involved in the induction of apoptosis in vivo, contributing to the rapid depletion of cells in the IBDV-infected bursa.

Animals↗

PubMed1995

Efficacy of subcutaneous application of live infectious bursal disease vaccine in young chickens with maternally derived antibody.

The infectivity of neutralized IBDV by normal chicken serum (NCS) was detected in day-old and 3-week-old chicken spleen adherent (CSA) cells, and that of neutralized IBDV by maternal antibody (MN-Ab) was detected in 3-week-old CSA cells. Moreover, CSA cells from day-old chickens had complement receptor (CR), and CSA cells from 1-week-old had both CR and Fc receptor (FcR). However, the infectivity of neutralized IBDV by MN-Ab was confirmed on CSA cells which were blocked for FcR on CSA cells by heat-aggregated NCS (56 degrees C, 60 min). These results indicated that infection of neutralized IBDV by NCS on CSA cells occurred via CR, and neutralized IBDV by MN-Ab was infected via FcR. In day-old specific pathogen free (SPF) chickens, the antibody level in NCS treated and non-treated IBD live vaccine subcutaneously inoculated groups was higher than the levels in the MN-Ab-treated IBDV inoculated group, and detected until 28 days old. Moreover, subcutaneously inoculated chickens were protected against the challenge of wild IBDV at 21 days old, whereas subcutaneously inoculated chickens were infected with MN-Ab-treated IBD live vaccine. In commercial layers which had MN-Ab, antibody levels of subcutaneously vaccinated group were higher than both the non-vaccinated and orally vaccinated groups, and virus isolation and viral antigen were positive with high detection rates on peripheral lymphocytes of each subcutaneously vaccinated group of SPF and commercial chickens.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

PubMed1999

Sequence comparison of the VP2 variable region of infectious bursal disease virus isolates from Vietnam.

The variable region in the VP2 gene of twenty-three infectious bursal disease virus (IBDV) isolates, collected in Vietnam in 1997 and 1998, was amplified as cDNA by using the reverse transcription-polymerase chain reaction and sequenced. Analysis of amino acid substitutions and phylogenetic relationships of the deduced amino acid sequences (residues 206-350) showed that the nineteen Vietnamese vv IBDVs clustered with the European vv IBDVs, Japanese vv IBDVs and Chinese vv strains, and that the four vietnamese virulent strains were closely related to European virulent strain 52/70. These results suggest that Vietnamese vv IBDVs, European vv IBDVs, Japanese vv IBDVs and Chinese vv strains have the same origin.

Amino Acid Sequence↗

PubMedDate not supplied

Immunosuppression in specific-pathogen-free broilers administered infectious bursal disease virus vaccines by in ovo route.

The effect of two infectious bursal disease virus (IBDV) vaccines (IBDV-immune complex [Icx] and IBDV-2512), administered in ovo, on the cell-mediated immunity of specific-pathogen-free (SPF) broilers was examined. A decrease (P < 0.05) in the T-cell mitogenic response occurred in birds vaccinated with both vaccines on days 9 and 21 post in ovo vaccination (PIOV), but an increase (P < 0.05) occurred on day 15 PIOV. The T cells from birds given the IBDV-2512 were less responsive. There were no significant differences in proportions of lymphocytes expressing CT4+CT8 and CT8+CT4- except on day 21 PIOV, where an increase (P < 0.05) in IBDV-2512-vaccinated birds and a decrease (P < 0.05) in percentage of CT4+CT8- in IBDV-Icx-vaccinated birds was observed. There was an increase (P < 0.05) in percentage of CT8+CT4- T cells on day 21 PIOV in both vaccinated groups. A decrease (P < 0.05) in B-cell percentage was observed on day 21 PIOV in birds given both vaccines. Results indicated that although humoral immunosuppression is associated with destruction of B cells (bursal atrophy), cell-mediated immunosuppression induced by these two IBDV vaccines in SPF birds was not associated with altered helper (CT4+CT8-) or cytotoxic (CT8+CT4-) subpopulations of T lymphocytes.

Animals↗

PubMedDate not supplied

Amino acid comparison of infectious bursal disease viruses placed in the same or different molecular groups by RT/PCR-RFLP.

Infectious bursal disease virus (IBDV) strains have been identified and placed into molecular groups by a reverse transcriptase (RT)/polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay. The predicted amino acid sequences corresponding to the region of the genome examined by RFLP were determined and compared for 14 IBDV strains from different molecular groups and 11 IBDV strains that were identified in molecular group 6. Among the viruses within molecular group 6, 13 amino acid positions had mutations, and among the viruses in different molecular groups, 27 amino acid positions had mutations. In addition to having more mutations, viruses compared from different molecular groups also had mutations at key positions that were previously reported to be important for the formation of neutralizing epitopes. Three of these IBDV strains with unique RFLP patterns were used to challenge 1-wk-old broiler chickens with maternal immunity to IBDV. One of these viruses, T1, broke through this maternal immunity as evidenced by detection of the virus by RT/PCR-RFLP and production of an active virus neutralizing antibody response to classic and variant IBDV strains. Unique amino acid mutations in the T1 virus that may have contributed to its ability to break through this maternal immunity were observed at amino acids 318 and 322. The results indicate that RFLP profiles and nucleotide sequences can be used to predict the relative similarities and differences among IBDV strains, but determining the actual antigenic differences among viruses requires testing in vivo.

Amino Acid Sequence↗

PubMed1992

Differential detection of infectious bursal disease virus serotypes, using cDNA probes to VP2 coding region.

Two nonoverlapping clones, pOH405 and pOH632, containing cDNA inserts in the VP2 coding region of genome segment A were selected from a cDNA library prepared from the double-stranded RNA genome of the OH strain of infectious bursal disease virus (IBDV) of serotype 2. Clone pOH405, which is located in the hypervariable segment of VP2, is 328 base pairs long, has nucleotide sequence homology of 72 to 73%, and amino acid sequence homology of 64 to 67% with IBDV strains of serotype 1. Clone pOH632, which is located in the highly conserved C-terminal part of VP2, is 230 base pairs long, has nucleotide sequence homology of 87 to 88%, and amino acid sequence homology of 100% with IBDV serotype 1. The lower detection limit of 32P-labeled probes prepared from both clones was 10 ng of OH-IBDV double-stranded RNA, using high-stringency conditions of hybridization (54 C, 50% formamide) and washing (55 C, 0.015M NaCl, 0.0015M trisodium citrate, pH 7.0, with 0.1% sodium dodecyl sulfate), and autoradiography for 24 hours. Under these conditions, the dot-blot hybridization assay for detection of serotype 2 IBDV double-stranded RNA, was 1,000 times more sensitive, using probe pOH632, but only 10 times more sensitive, using probe pOH405, compared with the assay for IBDV serotype 1, using the same probes. Thus, probe pOH632 could differentiate between the 2 IBDV serotypes by nucleic acid hybridization.

Amino Acid Sequence↗

PubMed2005

Molecular characterization of infectious bursal disease virus isolates from Nepal based on hypervariable region of VP2 gene.

Two Infectious bursal disease virus (IBDV) isolates, NP1SSH and NP2K were obtained from a severe infectious bursal disease (IBD) outbreak in Nepal in 2002. The hypervariable (HV) region of VP2 gene (1326 bp) of the isolates was generated by RT-PCR and sequenced. The obtained nucleotide sequences were compared with those of twenty other IBDV isolates/strains. Phylogenetic analysis based on this comparison revealed that NP1SSH and NP2K clustered with very virulent (vv) IBDV strains of serotype 1. In contrast, classical, Australian classical and attenuated strains of serotype 1 and avirulent IBDV strains of serotype 2 formed a different cluster. The deduced amino acid sequences of the two isolates showed a 98.3% identity with each other and 97.1% and 98.3% identities, respectively with very virulent IBDV (vvIBDV) isolates/strains. Three amino acids substitutions at positions 300 (E-->A), 308 (I-->F) and 334 (A-->P) within the HV region were common for both the isolates. The amino acids substitutions at positions 27 (S-->T), 28 (I-->T), 31 (D-->A), 36 (H-->Y), 135 (E-->G), 223 (G-->S), 225 (V-->I), 351 (L-->I), 352 (V-->E) and 399 (I-->S) for NP1SSH and at position 438 (I-->S) for NP2K were unique and differed from other IBDV isolates/strains. NP1SSH and NP2K showed highest similarity (97.8%) with the BD399 strain from Bangladesh as compared with other vvIBDV isolates/strains. We conclude that the NP1SSH and NP2K isolates of IBDV from Nepal represent vvIBDV of serotype 1.

Amino Acid Sequence↗

PubMedDate not supplied

Single and combined infections of specific-pathogen-free chickens with infectious bursal disease virus and an intestinal isolate of reovirus.

The susceptibility of 1-day-old and 7-day-old specific-pathogen-free chickens to infection with a virulent strain of infectious bursal disease virus (IBDV) or an intestinal isolate of avian reovirus, or a combination of the two, was investigated. Chickens infected with IBDV and reovirus had more severe pathological lesions than chickens infected with either virus alone, and prior infection with IBDV enhanced the pathogenicity of enteric reovirus. Virus recovery was attempted from bursa, spleen, thymus, liver, intestine, pancreas, cecal tonsils, heart, and tarso-metatarsal tendons. Viruses were recovered from all tissues sampled for either IBDV or reovirus isolation, and indications were that infection with IBDV before infection with reovirus led to longer persistence of reovirus in some tissues. Antibodies to IBDV or reovirus were measured by the virus neutralization test and enzyme-linked immunosorbent assay. Chickens infected with IBDV had lower (P less than 0.05) antibody responses to reovirus than chickens infected with reovirus alone.

Animals↗

PubMedDate not supplied

Effect of maternal antibody on timing of initial vaccination of young white leghorn chickens against infectious bursal disease virus.

Maternal antibody titers in white leghorn chicks against infectious bursal disease virus (IBDV) were measured by a computer-assisted, single-serum-dilution, indirect kinetic-based enzyme-linked immunosorbent assay (KELISA) and by a virus-neutralization (VN) test in order to predict the timing of initial vaccination. Day-old white leghorns were from unvaccinated pullets or from pullets vaccinated either four times or twice with IBDV commercial vaccines. The chicks were immunized once via the drinking water with a commercial "intermediate" live IBDV vaccine at 1, 15, or 28 days of age. Effective initial immunization was confirmed by an increase in antibody to IBDV (serologic conversion) that occurred when maternal antibody decreased to 8 and 9 on a log2 scale. This concentration of antibody was detected between 24 and 28 days of age. The computer-assisted IBDV-KELISA increased the sample processing speed for detecting IBDV antibody, and it was as sensitive as the VN test for predicting the timing of initial IBDV vaccination.

Animals↗

PubMedDate not supplied

Failure of two serotype II infectious bursal disease viruses to affect the humoral immune response of turkeys.

The effect of serotype II infectious bursal disease virus (IBDV) isolates from turkeys on the homoral immune response of turkey poults was determined. Following exposure to the OH IBDV isolate, poults in two experiments were inoculated with sheep red blood cells, which is a T-dependent antigen, and poults in two other experiments were inoculated with Salmonella heidelberg O antigen, which is a T-independent antigen. Prior exposure to serotype II IBDV did not affect serum antibody titers to these antigens. IBDV infection also did not affect the concentrations of serum immunoglobulin M (IgM), IgG, or IgA in these poults. Bursa:body weight ratios of OH IBDV-infected poults were not significantly different from those of uninfected controls. In one experiment, the humoral immune response of poults to the LaSota Newcastle disease vaccine was not affected by infection with the MO IBDV isolate. Although no clinical infectious bursal disease was observed in any poult in these experiments, the serotype II IBDV isolates were infectious and transmissible in poults.

Animals↗

PubMedDate not supplied

Plasma cell changes in the gland of Harder following infectious bursal disease virus infection of the chicken.

Tissue changes in the gland of Harder (GH) were studied following infection of day-old chicks with infectious bursal disease virus (IBDV). Broiler chicks that lacked maternal antibody to IBDV were inoculated with IBDV at 1 day of age and were maintained segregated from uninoculated controls. Plasma cell (PC) content of the GH was lowered among infected chickens from 1 to 7 weeks postinoculation. Since the GH in uninoculated control birds was not densely populated with PCs until 3 weeks of age, the PC depletion found in young chickens infected with IBDV became more clear with age. After the chickens were 3 weeks old, fewer (P less than .01) plasma cells were observed in the GH of IBDV-infected chicks than in uninoculated controls. Lymphoid follicles and heterophil populations in the GH did not appear to be affected by IBDV infection, nor could necrotic or degenerative changes be found in the acini or excretory ducts. The observation of fewer PCs suggests that a major lymphoid tissue of the upper respiratory tract is comprised by IBDV infection of day-old chicks.U

Animals↗

PubMedDate not supplied

Growth and infectivity titration of virulent infectious bursal disease virus in established cell lines from lymphoid leukosis tumors.

Lymphoblastoid cell lines were investigated for susceptibility to virulent (wild) infectious bursal disease virus (IBDV) and attenuated IBDV by detection of fluorescent antigen in inoculated cells. LSCC-BK3 and LSCC-CU10 cells were susceptible to both types of IBDV; LSCC-1104-B-1 line was susceptible to only the attenuated IBDV. However, 2 lymphoid leukosis and 2 Marek's disease cell lines were refractory to growth of any type of IBDV. When the J1 strain of virulent IBDV was inoculated into LSCC-BK3 cells, intracellular and extracellular viruses began to increase logarithmically between 4 and 8 hr postinoculation and reached 10(6.3) and 10(7.3) TCID50/0.1 ml, respectively, at 72 hr. Infectivity titration and serum-neutralization tests of virulent IBDV were possible with LSCC-BK3 cells.

Animals↗

PubMedDate not supplied

Maternal antibody and its effect on infectious bursal disease immunization.

Chickens vaccinated with infectious bursal disease virus (IBDV) early in life and revaccinated with an inactivated, oil-adjuvant IBDV vaccine at 18 weeks of age produced and maintained high levels of virus-neutralizing (VN) antibody through 10 months of lay. VN-antibody titers of chicks hatched from eggs laid during the same period closely matched the average VN-antibody titers of the dams. A sequential study of the decline rates of IBDV maternal antibody (MAB) in unvaccinated and IBDV-vaccinated chicks showed that the vaccine virus did not accelerate the antibody depletion rate in vaccinated chicks. Chicks carrying high IBDV MAB showed no active immune response to vaccination with commercial IBDV vaccines. They were also refractory to a pathogenic field isolate of IBDV (FV). However, chicks with low levels of MAB responded to both vaccine virus and the FV, although their response to vaccine virus was milder and delayed.

Animals↗