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PubMed2006

Pathogenicity of fowl adenovirus isolated from gizzard erosions to immuno-suppressed chickens.

Pathogenicity of a fowl adenovirus (FAV), JM1/1 strain of serotype 1 derived from gizzard erosions of a broiler chicken, was examined to specific pathogen-free (SPF) chickens pre-treated with infectious bursal disease viruses (IBDVs) or cyclophosphamide (CY). Virulent IBDVs, classical type, were inoculated orally at 3 days of age of SPF chickens. CY was treated subcutaneously for 3 days after hatch. FAV was given orally at 30 days of age. At 40 days of age, all chickens were bled and autopsied for serology and gross observation. Gizzard lesions were ranked by the scores depending on their severities. IBDV- or CY-treated chickens showed significantly higher gizzard lesion scores than non treated birds. There were no gross lesions in any other organs except for bursal atrophy. Serologically, antibody production against FAV was highly suppressed by IBDV infection or CY treatment.

Adenoviridae Infections↗

PubMed2000

Pathological and immunological study of an in ovo complex vaccine against infectious bursal disease.

The appearance of very virulent strains of infectious bursal disease (IBD) virus at the end of the 1980s made it necessary to develop more effective immunization procedures. To facilitate this, the immunogenicity and the immunosuppressive effect of a mild (G-87), an intermediate (LIBD) and an intermediate-plus (IBDV 2512) IBDV strain were tested after the in ovo inoculation of 18-day-old SPF and broiler chicken embryos. It was established that no noteworthy difference existed between the immunized and the control embryos in hatching rate and hatching weight. The higher the virulence of the vaccine virus strain, the more severe damage it caused to the lymphocytes of the bursa of Fabricius. In SPF chickens, the haemagglutination inhibition (HI) titres induced by a Newcastle disease (ND) vaccine administered at day old decreased in inverse ratio to the virulence of the IBD vaccine strain, while in broiler chickens this was not observed. Despite the decrease of the HI titre, the level of protection did not decline, or did so only after the use of the 'hot' strain. SPF chickens immunized in ovo with a complex vaccine prepared from strain IBDV 2512 and IBD antibody showed the same protection against Newcastle disease as the broilers. In broiler chicken embryos immunized in ovo, only strain IBDV 2512 induced antibody production, and such chickens were protected against IBD at 3 weeks of age. The complex vaccine administered in ovo has been used successfully at farm hatcheries as well.

Animals↗

PubMed1998

Western blot detection of infectious bursal disease virus infection.

In order to evaluate the use of a Western blot methodology for the diagnosis of infectious bursal disease virus (IBDV) infection, chickens were experimentally infected with IBDV strains and tested for the presence of viral antigens and antibodies by a blocking Western blot test (bWB). The viral proteins obtained from the bursa of Fabricius (BF) were transferred to a nitrocellulose membrane after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the chicken sera obtained by heart puncture were used for the detection of these proteins. In order to eliminate nonspecific reactions, we used a rabbit anti-chicken serum (blocking tool). By the use of the bWB test, two distinct viral proteins of 43-kDa (VP2) and 32-kDa (VP3) were detected. We suggest the use of this methodology for the detection of IBDV infection in animals suspected of having IBDV reinfection and a chronic subclinical form of the disease. With the use of the rabbit anti-chicken sera for blocking, this method is practical, sensitive and less time consuming.

Animals↗

PubMed2006

Inactivation of infectious bursal disease virus by binary ethylenimine and formalin.

In this experiment conducted to study the inactivation dynamics of infectious bursal disease virus (IBDV) by binary ethylenimine (BEI) in comparison with formalin, IBDV was isolated from the bursa of infected chickens and its confirmation was done by agar gel precipitation test. Viral suspensions were subjected to inactivation with BEI and formalin for pre-set time intervals. BEI was employed at concentrations of 0.001 and 0.002 mol/L while formalin was used at 0.1% and 0.2%. Sampling was done at 6, 12, 24, 36 and 48 h of incubation and samples were tested for their inactivation status in 9-day-old embryonated eggs and 3-week-old broiler chickens. IBDV was completely inactivated by 0.001 and 0.002 mol/L BEI after 36 h of incubation at 37 degrees C, whereas formalin at 0.1% and 0.2% concentrations inactivated IBDV in 24 h.

Animals↗

PubMedDate not supplied

Pathogenesis and tissue distribution of a variant strain of infectious bursal disease virus in commercial broiler chickens.

The detection of either infectious virus, viral antigen, and/or viral RNA in different tissues of commercial broilers inoculated at 1 day of age with E/Del variant strain of infectious bursal disease virus (IBDV) was investigated at 2, 4, and 6 wk postinoculation (PI). Virus was readily isolated from homogenates of bursa, cecal tonsils, and bone marrow at 2 and 4 wk PI. Virus isolation coupled with immunoperoxidase assay or reverse transcription-polymerase chain reaction for IBDV-specific RNA extended the window of IBDV detection in the bursa of Fabricius to 6 wk PI. Serology indicated an active early virus infection; however, viral pathology was observed later and beginning at 4 wk PI. This study indicates that variant strains of IBDV may be present in commercial broilers longer than previously thought, and cecal tonsils and bone marrow may serve as nonbursal lymphoid tissues supporting virus replication at later time points PI.

Animals↗

PubMedDate not supplied

Differentiation of infectious bursal disease viruses isolated in Croatia.

Infectious bursal disease viruses (IBDVs) in 26 IBDV-positive bursa samples collected in Croatia during the period 1996-2000 and in two commercially available vaccines were differentiated by the presence or absence of the CfoI, SacI, SspI, StuI, and TaqI restriction sites in the 422-bp fragment of segment A of the VP2 gene (nt 732-1153). The fragments from 14 (54%) field isolates were TaqI+ StuI+ SspI+ and SacI- CfoI-, indicating their very virulent (vv) character. The presence of CfoI restriction site in 10 (38%) field isolates is uncommon for vvIBDV strains. It was detected in only the 88180 vvIBDV strain. Nevertheless, these isolates can be classified as vv strains according to TaqI+ StuI+ SspI+ SacI- restrictions. Two SacI+ StuI+ CfoI+ TaqI- SspI- field isolates (8%) could be classified as non-vvIBDVs. The StuI+ restriction is common to vvIBDV strains. However, the StuI recognition sequence is present in the F52/70 classic European and 002-73 attenuated strains as well. The SacI+ CfoI+ StuI- SspI- restrictions and the lack of the TaqI restriction at nt position 832 show that the IBDV in GUMBOKAL IM-SPF vaccine corresponds to the attenuated and/or vaccine strains. The TaqI restriction at nt position 875 suggests that the IBDV in GUMBOKAL SPF vaccine could belong to the mild strains.

Animals↗

PubMedDate not supplied

Sequence analysis of Malaysian infectious bursal disease virus isolate and the use of reverse transcriptase nested polymerase chain reaction enzyme-linked immunosorbent assay for the detection of VP2 hypervariable region.

The VP2 hypervariable region of P97/302 local infectious bursal disease virus (IBDV) isolate was amplified by the reverse transcriptase (RT) nested polymerase chain reaction (PCR) and cloned. This region of P97/302 local isolate was sequenced and compared with eight other reported IBDV sequences. The result showed that P97/302 IBDV was most identical to the reported very virulent IBDV strains because it has amino acid substitutions at positions 222, 256, 294, and 299, which encode alanine, isoleucine, isoleucine, and serine, respectively. This region can be digested with restriction enzymes of Taq1, Sty1, Ssp1 but not with Sac1. The P97/302 isolate was then used for the optimization of RT nested PCR enzyme-linked immunosorbent assay (ELISA). The RT nested PCR ELISA was able to detect 10(-4) dilution of the infected bursa homogenates and was 10 times more sensitive when compared with the agarose gel detection method. The RT nested PCR ELISA can detect up to 0.48 ng of the PCR product. The specificity of this nested PCR ELISA was also high (100%).

Amino Acid Sequence↗

PubMedDate not supplied

Very virulent infectious bursal disease virus in southeastern Europe.

Clinical outbreaks of severe acute infectious burial disease (IBD) were recorded since the mid- and late 1990s in several countries in the southeastern part of Europe. Epidemiologic data showed that both infectious bursal disease virus (IBDV)-vaccinated and IBDV-nonvaccinated chickens were affected with acute IBD and mortality up to 50% independent of the IBDV vaccination status of the appropriate parent flocks. For investigation of the causative agent of acute IBD, the variable region of VP2 was amplified, cloned, and sequenced. Nucleotide sequence analysis of polymerase chain reaction fragments showed several silent nucleotide exchanges in comparison with the sequence of the very virulent (vv) IBDV strain UK661. Also, restriction enzyme cleavage sites proposed specific for vvIBDV were present in all investigated strains. On the basis of clinical signs in affected flocks, recorded epidemiologic data, and sequence analysis, it is very likely the IBD-causing strains were of the vv phenotype.

Aging↗

PubMedDate not supplied

Characterization of Egyptian field strains of infectious bursal disease virus.

The reverse transcriptase-polymerase chain reaction/restriction fragment length polymorphism (RT-PCR/RFLP) technique was used for identification and characterization of Egyptian field strains of infectious bursal disease viruses (IBDVs) that caused severe outbreaks with (30%-60%) mortality rate. Twenty-four bursal samples collected from 24 field outbreaks in commercially reared chicken flocks experiencing signs typical of infectious bursal disease (IBD) were used. Ten of the bursal samples examined were determined to contain IBDV as evidenced by amplification of a 743-bp region of the VP2 gene of IBDV by RT-PCR. The RT-PCR products of the detected viruses were characterized by digestion with three restriction enzymes, BstN, MboI, and SpI. Three different RT-PCR/RFLP profiles were observed. Seven of the detected viruses had RFLP profiles identical to the very virulent European strains of IBDV (vvIBDVs). One virus had a RFLP profile identical to the U.S. classic vaccine strain, and one virus had a unique RFLP profile. The clinical history of the outbreaks and the presence of the SspI site in the 743-bp RT-PCR fragment were the criteria for designating the viruses as belonging to the very virulent (vv) phenotype.

Base Sequence↗

PubMedDate not supplied

Pathogenicity and molecular analysis of an infectious bursal disease virus isolated from Malaysian village chickens.

The characteristics of the pathogenic infectious bursal disease virus (IBDV) that infected avian species other than commercial chickens were largely unknown. In this study, by using in vivo and molecular methods, we had characterized an IBDV isolate (named 94268) isolated from an infectious bursal disease (IBD) outbreak in Malaysian village chickens--the adulterated descendant of the Southeast Asian jungle fowl (Gallus bankiva) that were commonly reared in the backyard. The 94268 isolate was grouped as the very virulent IBDV (vvIBDV) strain because it caused severe lesions and a high mortality rate in village chickens (>88%) and experimentally infected specific-pathogen-free chickens (>66%). In addition, it possessed all of the vvIBDV molecular markers in its VP2 gene. Phylogenetic analysis using distance, maximum parsimony, and maximum likelihood methods revealed that 94268 was monophyletic with other vvIBDV isolates and closely related to the Malaysian vvIBDV isolates. Given that the VP2 gene of 94268 isolate was almost identical and evolutionarily closely related to other field IBDV isolates that affected the commercial chickens, we therefore concluded that IBD infections had spread across the farm boundary. IBD infection in the village chicken may represent an important part of the IBD epidemiology because these birds could harbor the vvIBDV strain and should not be overlooked in the control and prevention of the disease.

Animals↗

PubMed1983

Multivalent inactivated virus oil emulsion vaccines in broiler breeder chickens. II. Trivalent vaccines in breeders not previously vaccinated with live Newcastle disease, infectious bursal disease, and tenosynovitis vaccines.

Inactivated Newcastle disease (NDV), infectious bursal disease (IBDV), and viral arthritis/tenosynovitis (VA) viruses were incorporated into water-in-oil emulsion vaccines either alone, in bivalent combinations, or in a trivalent vaccine. Twenty-week-old broiler breeder chickens with no previous exposure to NDV, IBDV, or VA live virus vaccines were injected intramuscularly with the monovalent, bivalent, or trivalent vaccines. The antibody responses to NDV in all three vaccines were poor, and NDV-hemagglutination-inhibition (HI) geometric mean titers (GMTs) never rose above 20 during the 40-week trial. The antibody response to IBDV showed a strong primary response 4 weeks after vaccination, but IBD-VN geometric mean titers declined steadily to less than 100 by 4 months after vaccination. The antibody GMTs to IBDV continued to decline for the remainder of the trial. The antibody response to VA virus was biphasic, with peak VA-virus neutralization (VN) geometric mean titers occurring at 3 months and 6 months postvaccination. The amplitude of the response to the monovalent, bivalent, and trivalent vaccines was inversely proportional to the number of antigens incorporated into each vaccine. Maternal antibody titers in the progeny against each of the three antigens reflected those of the parents. In no case were maternal antibody titers detectable beyond 14 days of age.

Animals↗

PubMed1984

Vaccination against Marek's disease and infectious bursal disease. II. Titration and in vivo efficacy of the coinfection-derived bivalent live vaccine for turkey herpesvirus and infectious bursal disease virus.

The bivalent HVT/IBDV live vaccine, developed by cocultivation of the turkey herpesvirus (HVT) and infectious bursal disease virus (IBDV) vaccine in cell cultures, was quantitated. The titer of each constituent virus could be successfully determined in vitro by pretreatment of the vaccine with anti-IBDV serum or chloroform (CHCl3). Studies in vivo further demonstrated that the anti-IBDV serum-treated HVT/IBDV vaccine lost its immunogenicity by failing to induce IBDV-neutralizing antibody or to protect vaccinates from bursal atrophy after challenge exposure to infectious bursal disease. The CHCl3 treatment significantly reduced the efficacy of the vaccine to protect vaccinates against Marek's disease challenge.

Animals↗

PubMedDate not supplied

Organ culture of chicken bursa as a model to study the pathogenicity of infectious bursal disease virus isolates.

In vitro study with chicken bursal organ culture was attempted to assess the pathogenicity of locally isolated infectious bursal disease virus (IBDV) initially isolated from the bursa of naturally infected birds. In bursal organ culture, lymphoblastic transformation was noticed as early as 24 hr postinoculation and reached maximum at 72 hr postinoculation. The other microscopic changes were increased number of macrophages and formation of plasma cells. The IBDV antigen was detected 24 hr onward by coagglutination test with antibody coated Staphylococcus aureus strain Cowan I. On the basis of lesion score, the three isolates of IBDV (A, B, and C) were graded as virulent (B isolate) and moderately virulent (A and C isolates). A similar pattern of pathogenicity was also observed in the in vivo pathogenicity studies in chicken based on bursa: body weight ratio and histopathologic lesion score. The bursal organ culture thus provides a useful experimental model to differentiate the IBDV isolates on the basis of their virulence.

Animals↗

PubMedDate not supplied

Correlation of enzyme-linked immunosorbent assay titers with protection against infectious bursal disease virus.

We examined the utility of baculovirus-expressed infectious bursal disease virus (IBDV) proteins to act as antigens in the enzyme-linked immunosorbent assay (ELISA). The three IBDV protein antigens tested included 1) a truncated VP2, 2) whole VP2, and 3) the polyprotein products VP2, VP3, and VP4. Serum samples from 2-wk-old commercially reared broilers were collected and tested in the three ELISAs. Serum samples were obtained from 34 different commercial broiler flocks. An average of 14 serum samples (range = 11-17) were tested for each flock. The ELISA results were compared with the percentage of protection of these birds following challenge with IBDV. Fifty 2-wk-old chicks from each of the 34 broiler flocks were challenged with STC classic virus or Del-E variant virus. At 7 days postchallenge, the bursa from each of the birds was removed and bursa/body weights were recorded. Percentage of protection was determined by the number of birds in each challenge group that had normal relative bursal weights compared with unchallenged controls. No evidence was found of a relationship between ELISA data generated with the polyprotein antigen (VP2, VP3, VP4) and percentage of protection observed in the STC and Del-E challenged birds. A significant relationship was found between ELISA data and percentage of protection to STC and Del-E when the truncated VP2 or whole VP2 antigens were used in the ELISA. The results of this study indicate that predicting the percentage of protection against classic or variant IBDV strains in broilers from vaccinated breeder flocks can be improved when VP2 is used as the only antigen in the ELISA.

Animals↗

PubMedDate not supplied

Antigenic and molecular characterization of three infectious bursal disease virus field isolates.

Three infectious bursal disease field viruses, identified as U28, 3212, and MISS and isolated in the early 1980s from the southeastern United States, were characterized both antigenically and genotypically. A panel of monoclonal antibodies (MAbs) was utilized in an antigen-capture enzyme-linked immunosorbent assay (ELISA) for antigenic characterization. The ELISA data indicated that U28 and the Delaware variants are antigenically related. The 3212 and the GLS variants were more closely related antigenically to each other than to other viruses analyzed. However, the MISS isolate reacted with MAbs that were specific for both classic and variant strains of infectious bursal disease virus (IBDV). Reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify nucleotide sequences from the genome coding for the variable region of VP2 from IBDV field isolates U28, 3212, and MISS. Phylogenetic analysis of the deduced amino acid sequences revealed that U28 was most similar (98.3%) to the IBDV Delaware variants and that 3212 was most similar (97.1%) to the GLS variant. The MISS isolate was most similar (97.4%) to the classic 52/70 strain. Positive correlation occurred with the use of different methods to characterize IBDV isolates.

Animals↗

PubMedDate not supplied

In ovo vaccination of specific-pathogen-free chickens with vaccines containing multiple agents.

We used in ovo technology to protect chickens against multiple diseases by inoculating vaccines containing mixtures of live viral agents. A single in ovo injection of a vaccine containing serotypes 1, 2, and 3 of Marek's disease virus (MDV), a vaccine strain of serotype 1 infectious bursal disease virus (IBDV), and recombinant fowl pox vaccine with HN and F genes of Newcastle disease virus (rFP-NDV) induced protection against virulent MDV, IBDV, Newcastle disease virus, and fowl poxvirus. The multiple-agent vaccine induced specific antibodies against the viral agents present in the mixture and did not adversely affect the survival of hatched chickens. Inoculation of a vaccine containing serotypes 1, 2, and 3 of MDV and IBDV did not affect hatchability of eggs, although the addition of rFP-NDV to the mixture reduced hatchability by 23%-26%. In ovo vaccination with a vaccine containing MDV and IBDV vaccine viruses did not exacerbate the inhibitory effect of individual viral agents on humoral and cellular immune competence.

Animals↗

PubMedDate not supplied

Restriction fragment length polymorphisms in the VP2 gene of infectious bursal disease viruses from outside the United States.

Infectious bursal disease viruses (IBDVs) were identified in bursa samples from chickens reared outside the United States. All samples were obtained from commercially reared chicken flocks experiencing signs typical of infectious bursal disease. The reverse transcriptase/polymerase chain reaction (RT/PCR) technique was used to detect the viruses and restriction fragment length polymorphism (RFLP) was used to compare a 743-bp region of the VP2 gene among the viruses detected. The 81 IBDVs detected were from 16 different countries and were assigned to molecular groups on the basis of their RFLP following digestion with the restriction enzymes BstNI and MboI. The presence of an SspI site in the 743-bp RT/PCR fragment was used to predict the very virulent (vv) phenotype. Almost half (49%) of the viruses detected had the same RFLP with the BstNI and MboI enzymes. These viruses were placed into a new molecular group, designated group 6, that was exemplified by the RS593 vaccine strain of IBDV, which also had this molecular RFLP pattern. Some viruses detected had RFLP patterns similar to molecular groups 2, 3, and 4, previously described in our laboratory. Sixteen RFLP patterns were observed that did not match RFLP results we previously obtained for vaccine IBDV strains. The SspI restriction site was found in 46% of the viruses detected, predicting that these viruses have the vvIBDV phenotype. The SspI site was not observed in viruses from Central and South America.

Animal Husbandry↗

PubMedDate not supplied

Experimental inoculation of avian polyomavirus in chemically and virally immunosuppressed chickens.

The purpose of this series of experiments was to determine the effect of various types of immunosuppressive treatments (cyclophosphamide, infectious bursal disease virus [IBDV], chicken anemia virus [CAV], and combination infection with IBDV and CAV) on susceptibility of chickens to challenge with avian polyomavirus. In the first experiment, chickens were chemically bursectomized with intraperitoneal injections of cyclophosphamide; in the second study, chickens were orally inoculated with IBDV; in the third study, birds were intramuscularly inoculated with CAV; and in the final study, birds were inoculated with both IBDV and CAV. In all experiments, chickens were challenged with 10(4.7) tissue culture infective doses of polyomavirus intraperitoneally. Only chemically bursectomized chickens developed lesions similar to those found in the naturally occurring multisystemic fatal form of polyomavirus infection seen in psittacine nestlings, including hepatic necrosis and large pale intranuclear inclusions.

Animals↗