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1.
Two infectious bursal disease viruses (IBDVs 1174 and V1) were isolated from IBDV-vaccinated broiler flocks in California and Georgia. These flocks had a history of subclinical immunosuppression. These isolates are commonly used in IBDV progeny challenge studies at Auburn, AL, as well as vaccine manufacturer's vaccine efficacy studies, because they come from populated poultry-producing states, and are requested by poultry veterinarians from those states. Nested polymerase chain reaction (PCR) generated viral genome products for sequencing. A 491-bp segment from the VP2 gene, covering the hypervariable region, from each isolate was analyzed and compared with previously sequenced isolates. Sequence analysis showed that they were more closely related to the Delaware (Del) E antigenic variant than they are to the Animal Health Plant Inspection Service (APHIS) standard, both at the nucleotide level (96%, 97%) and at the amino acid level (94%, 97%). Both isolates had the glutamine to lysine shift in amino acid 249 which has been reported to be critical in binding the virus neutralizing Mab B69. Phenotypic studies showed that both isolates produced rapid atrophy of the bursae and weight loss, without the edematous bursal phase, in 2-wk-old commercial broilers having antibody against IBDV. A progeny challenge study showed both isolates produced more atrophy of the bursae (less percentage of protection) than the Del E isolate. Molecular and phenotypic data of these important IBDV isolates help in the improved detection and control of this continually changing and important viral pathogen of chickens.  相似文献   

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OBJECTIVE: To characterise infectious bursal disease viruses (IBDVs) prevalent at major commercial sites throughout Australia and to compare the nucleic acid sequences of local strains of IBDV with those of characterised overseas strains. DESIGN: Samples of bursae were collected from 20 broiler farms that belonged to different poultry companies in New South Wales (NSW), Queensland (Qld), Victoria (Vic), Westem (WA) and South Australia (SA). METHOD: Bursae were collected from broilers between 24 and 35 days of age. Bursal tissue was homogenised and tested for the presence of IBDV antigen using four monoclonal antibodies (Mabs) which detect antigenic variation in IBDV strains. The nucleotide sequences of the hypervariable region (HVR) within the VP2 gene of IBDVs was determined and the deduced amino acid sequences compared with three vaccine strains and six previously characterised Australian IBDV strains. The deduced amino acid sequences were also compared with the published amino acid sequences of overseas strains. The phylogenetic relationships between Australian strains and overseas strains were then determined. RESULTS: IBDV was detected in birds from 14 out of 20 farms sampled. Typing with four Mabs showed that all viruses from Vic (6) and SA (10) were antigenic variants, whereas all viruses from NSW (29), Qld (4) and WA (5) were classical-like strains. Nucleotide sequencing of one sample from each of the 14 farms on which IBDV was detected confirmed results obtained with Mabs. The amino acid sequences of all Australian viruses differed from the amino acid sequences of foreign IBDV strains. Phylogenetic analysis showed that Australian IBDV viruses belonged to two distinct genetic groups. Very virulent (vv) IBDV strains belonged to a third genetic group, and overseas classical and variant strains belonged to a fourth genetic group. CONCLUSIONS: The results confirmed previous findings that there are two groups of IBDV strains circulating in commercial broilers in Australia. The majority are classical-like strains that are antigenically and genetically similar to vaccine strains 002/73 and V877. These classical strains were prevalent in broilers in three states, NSW, Qld and WA. The second group of strains are antigenic variants that were only found in broilers in two states, Vic and SA. All Australian IBDVs characterised to date are genetically distinct and can be differentiated from all other overseas strains. This enables identification of incursion of any exotic strain into Australian poultry, be it classical, US variant or wIBDV strains.  相似文献   

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A field study was designed to determine the prevalence of subclinical infectious bursal disease (IBD) in broiler chickens from a commercial poultry company. Bursae of Fabricius (BF) from two vaccinated and three nonvaccinated broiler flocks were evaluated histologically, and antibody profiles of these broiler and matched parent breeder flocks were established. Lesions of IBD, including lymphoid necrosis, stromal edema, and infiltrates of heterophils and macrophages, were first detected in BF at 24 days of age in both vaccinated and nonvaccinated chickens. At 41 days, all BF had lesions characteristic of IBD, including severe lymphoid depletion, proliferation of epithelial cells, and mild fibroplasia. Although mean maternal antibody levels (measured by enzyme-linked immunosorbent assay) in broilers were apparently protective through day 12, IBD antibodies decreased to nonprotective levels (below 1,000) by day 16 or 20. Titers began to increase by day 28 or 32 because of field exposure. Sentinel birds, placed with broiler flocks, also developed IBD antibody titers. Broiler breeders had low and nonuniform antibody titers. Prevalence of field IBD exposure was high, and existing vaccination programs were not effective.  相似文献   

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Using a sentinel bird approach, two field isolates of infectious bursal disease virus (IBDV) were isolated from broiler farms in two major broiler-producing areas of the state of Georgia. These farms had a history of subclinical IBD associated with respiratory problems and poor performance. Isolates designated as U-28 and 3212 were isolated using specific-pathogen-free chicken embryos and chicken embryo bursal cells. These isolates were identified by means of agar gel precipitation and virus-neutralization tests, direct immunofluorescence, histopathology, and electron microscopy. Isolates U-28 and 3212 appear to differ in antigenicity and pathogenicity from previously known serotype I IBDV isolates. In evaluating the extent of bursal damage caused by these field isolates, an association was found between the bursa of Fabricius/body weight index, histopathology scoring of atrophy, and morphometric analysis of the total follicle area.  相似文献   

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In ovo vaccination against Marek's disease virus and infectious bursal disease virus (IBDV) in commercial broilers in the United States is common. Little information exists as to the safety and efficacy of intermediate IBDV vaccines given in ovo. Experiments were initiated to determine the safety and efficacy of three commercially available live intermediate IBDV vaccines by in ovo route. Commonly used vaccines were given at 18 days of embryonation to specific-pathogen-free (SPF) broiler embryos (first and second study) or to commercial broiler embryos (third study) that had maternal antibody against IBDV. When any of the antigenic standard vaccines was given at full dose to SPF embryos, embryonic and 3-wk posthatch mortality increased. Vaccines also caused significant microscopic lesions in the bursa of Fabricius at 1 and 3 wk posthatch. In contrast, there was no adverse effect on embryonic or posthatch mortality when vaccines were given at half dose to SPF or commercial broiler embryos. However, significant microscopic lesions were evident at 1 and 3 wk posthatch in the bursae of SPF embryos given the vaccines at half dose. When vaccines were given at half dose to commercial broiler embryos, lesions were evident at 1 but not 3 wk of age. In the third study, in ovo vaccinated chickens were challenged with either a virulent standard (APHIS) or antigenic variant (variant E) IBDV virus at 3 wk of age. All vaccines produced at least 87% protection against the standard and 60% protection against the variant challenge IBDV, as measured by bursal weight to body weight ratios. This study was the first to examine the safety and efficacy of the three commonly used intermediate IBDV vaccines given in ovo in protection against standard and antigenic variant IBDV challenge viruses.  相似文献   

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Experiments determined the efficacy of live vaccines in specific-pathogen-free broilers against serologic subtypes of infectious bursal disease virus (IBDV). Challenge isolates were the Delmarva variant E, a standard serotype I (APHIS) and a variant isolate from Mississippi. The vaccines were a cloned standard (CS) vaccine (Clone Vac-D78), a cloned variant (CV) vaccine (Bursa Vac IV), and an uncloned standard (UCS) vaccine (Bursine II). The severity of microscopic lesions was correlated with bursal atrophy as measured by bursa-weight-to-body-weight ratios. All vaccines provided adequate protection against the APHIS challenge. The three vaccines averaged 77% protection against APHIS in the first experiment and 78% in the second. Protection against the variant E and Miss isolates was considerably less for all vaccines. The three vaccines produced an average 70% protection against the Miss isolate in the first experiment and 69% in the second experiment. Against the variant E virus, the three vaccines averaged 67% protection in the first experiment and 65% in the second. There were significant differences in protection for each vaccine against individual IBDV subtypes. Results showed that no vaccine provided good protection (at least 80%) against all three subtypes of IBDV.  相似文献   

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Transgenic plants represent a safe, effective, and inexpensive way to produce vaccines. The immunogenicity of VP2 protein of an infectious bursal disease (IBD) virus variant E isolate expressed in transgenic Arabidopsis thaliana was compared with a commercial vaccine in specific-pathogen-free broiler chickens. The VP2 coding sequence was isolated and integrated into A. thaliana genome by Agrobacterium tumefaciens-mediated transformation. Soluble VP2 expressed in transgenic plants was used to immunize chickens. Chickens receiving oral immunization with plant-derived VP2 at 1 and 3 wk of age had an antibody response using enzyme-linked immunosorbent assay and 80% protection against challenge infection at 4 wk. Chickens primed with a commercial vaccine at 1 wk followed by an oral booster with VP2 expressed in plants at 3 wk of age showed 90% protection. Chickens immunized with a commercial vaccine at 1 and 3 wk showed 78% protection. Results supported the efficacy of plant-produced VP2 as a vaccine against IBD.  相似文献   

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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.  相似文献   

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An infectious bursal disease (IBD) outbreak occurred in the east region of Spain in the spring of 2002 and rapidly spread thorough the whole country, although proper vaccination programs were applied. In this report, 33 infectious bursal disease viruses (IBDVs) isolated from this outbreak were characterized by nucleotide sequencing of the VP2 gene hypervariable region and were compared with reference IBD strains and the 1990s Spanish IBDVs in order to determine possible emergence of IBDV isolates with modified antigenic or virulent properties. Moreover, histopathologic and immunohistochemical studies of those cases where bursal tissues were available were carried out. Of the 33 isolates, 23 were identified as very virulent IBDVs (vvIBDVs), whereas the other 10 isolates were classified as attenuated or intermediate virulence classical strains and could possibly be IBDV live vaccine strains used in the immunization of these chickens. Results of this study indicate that wIBDV isolates from the 2002 Spanish outbreak are closely related with those from the 1990s outbreak. However, acute IBD cases have not been reported in Spain during these 10 yr. Genetic, management, and environmental factors likely related with IBD reemergence in Spain are discussed. Moreover, our results indicate that good correlation exists between the IBDV subtype present in the field and the degree of lesions in bursa tissue, as well as the immunohistochemistry staining.  相似文献   

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A double-antibody sandwich ELISA was employed for detection of IBD virus in bursal suspension. Fifty IBD-free white leghorn chickens aged 5 weeks were experimentally infected with IBD virus. Bursae were collected 4, 8, 12, 24, 36 and 48 hours and 3, 4 and 5 days post-infection. An equal number of chickens acted as appropriate controls. The colour difference between a positive and a negative reaction was clearly distinguished with the naked eye. The cut-off level between ELISA negative and ELISA positive absorbance values was estimated at mean absorbance of negative controls plus three times the standard deviation.  相似文献   

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用RT-PCR方法从传染性法氏囊病(IBD)免疫预防失败的病鸡法氏囊组织AH1与AH2中扩增传染性法氏囊病病毒(IBDV)VP2基因。序列分析结果显示,AH1与AH2病毒VP2基因长度均为1350nt,编码450aa,核苷酸和氨基酸序列的同源性分别为98.2%、99.3%,七肽基序均为SWSASGS,在222、253、256、279、284、294和299位上的氨基酸残基分别是A、Q、I、D、A、I和S,具有IBDV强毒的分子特征。进一步将VP2基因克隆入人5型腺病毒穿梭载体(pShuttle-CMV),与腺病毒骨架载体(pAdEasyTM)共转化大肠杆菌BJ5183进行同源重组并转染HEK-293A细胞,经多次亚克隆获得了重组腺病毒rAd-(IBDV)VP2。利用Western-blot、IFA等方法检测IBDVVP2蛋白的体外表达情况,结果证明VP2基因在腺病毒中获得了表达。  相似文献   

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Infectious bursal disease virus variant from commercial Leghorn pullets   总被引:5,自引:0,他引:5  
An infectious bursal disease virus (IBDV) was isolated from 39-to-43-day-old commercial leghorn pullets suspected of having infectious bursal disease (IBD). These chickens had been vaccinated with a commercial live IBDV vaccine at 28 and 35 days of age. An isolate designated IN was recovered using specific-pathogen-free (SPF) chickens and the BGM-70 established cell line. Experimental studies using SPF chickens vaccinated with either inactivated vaccines made from the vaccine strain used in the problem flock or a standard-type vaccine indicated no protection against the IN isolate. However, two variants and another standard-type vaccine induced protection against the IN isolate. Cross-neutralization tests indicated that the IN isolate differed antigenically from commercial vaccine strains and was related to the variant IBDV strains recently isolated from broilers. To our knowledge, this is the first report of a variant IBDV recovered from commercial layer chickens in the United States.  相似文献   

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The effect of infectious bursal disease virus (IBDV) on Salmonella typhimurium (ST) infections in broilers was investigated in terms of Salmonella shedding and persistence, pathogenicity, and isotype specific humoral immune responses. Thirty-six, 1-day-old, straight-run commercial broiler chickens that were Salmonella negative by polymerase chain reaction (PCR) and culture were divided into two groups of 18 chicks each (ST and ST-IBDV). One group (ST-IBDV) of chicks received the E/Del strain of IBDV (10(5.0) median tissue culture infective dose [TCID50]/ml) through the ocular and cloacal routes divided into doses of 50 microl each at 2 days of age. Both groups were then inoculated with 10(8) colony-forming units (CFU)/ml nalidixic acid-resistant ST in the drinking water at 3 days of age. Environmental Salmonella counts were higher in the ST-IBDV group at 2 and 3 wk postinfection (PI) compared to the ST group. ST carriage in the cecal contents between the ST and ST-IBDV groups was not statistically different. The ST-IBDV group had a single mortality at 10 days postinfection compared to none in the ST group. The ST-IBDV group had significantly lower bursa to body weight ratios at 4 and 6 wk, as well as higher bursal lesion scores than the ST group at 2, 4, and 6 wk PI. The ST group had significant increase in serum IgG from 2 to 6 wk PI in comparison to the ST-IBDV group, which had no significant changes over time. Both IgA and IgM were significantly increased at 4 and 6 wk relative to 2-wk levels. There was an IBDV-induced failure of anti-Salmonella IgG seroconversion over time in ST-IBDV. Both groups continued to shed high levels of Salmonella up to the end of the study despite high antibody levels in the ST group and an unimpaired IgM and IgA production in the ST-IBDV group, indicating a limited influence of humoral immunity on Salmonella clearance.  相似文献   

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