Comparison of egg-yolk and serum antibody titers to four avian viruses by enzyme-linked immunosorbent assay using paired field samples

Eggs and blood were collected from 11 hens in each of nine broiler-breeder flocks in Quebec. Serum and egg-yolk extracts were assayed for antibody titers to infectious bursal disease virus (IBDV), infectious bronchitis virus (IBV), Newcastle disease virus (NDV), and reovirus (RV) by a commercial enzyme-linked immunosorbent assay (ELISA) kit. Comparison was made between egg-yolk and serum antibody titers by a regression analysis. A high correlation was observed between serum and yolk antibody titers to all the viruses tested (r = 0.9 for IBDV, 0.84 for IBV, 0.84 for NDV, and 0.91 for RV). Antibody monitoring of commercial breeder flocks using egg yolk instead of serum with commercial ELISA plates is thus feasible and is recommended.     

Rapid serological profiling by enzyme-linked immunosorbent assay. IV. Association of infectious bursal disease serology with broiler flock performance

Breeder and broiler flocks were serologically evaluated using a multiple enzyme-linked immunosorbent assay (M-ELISA). The serologic status of two commercial broiler-breeder flocks and their progeny was monitored, and 840 sera were promptly assessed for antibodies against six infectious agents using the M-ELISA. Breeder flocks were sampled at lay, and broiler chicks were hatched from fertile eggs collected on the scheduled lay date of the breeders. The broiler chicks were placed for growout as eight separate flocks (four from each breeder), and the serologic survey of broilers included sequentially sampling each flock five times between 1 day of age and market. Association of broiler vaccination schedules, mortality, and condemnation data with the temporal serologic data obtained indicated that the earlier appearance of active antibody against infectious bursal disease (IBD) in some unvaccinated flocks was associated with subsequent higher growout mortality and with the poorer overall performance that these flocks experienced. The results of this serologic survey also demonstrated that if a constant, well-timed monitoring program had not been used, major serologic differences between flocks would not have been detected. Serologic profiles of selected broiler flocks by virus-neutralization (VN) tests for infectious bronchitis virus (IBV) and reovirus or by hemagglutination-inhibition (HI) tests for Newcastle disease virus (NDV) compared favorably with the serologic profiles obtained by M-ELISA. Comparison of vaccination histories with serologic results derived from M-ELISA, VN or HI tests indicated that response to vaccination for IBV and NDV at 1 day was either blocked or significantly delayed by moderate levels of maternal antibody and/or were suppressed by an apparent field outbreak of IBD that occurred in all eight broiler flocks.     

Serological profiles of commercial broiler breeders and their progeny. 1. Infectious bronchitis virus

Serum samples were collected from broiler breeders and their 1-day-old, 2-week-old, and 5-week-old progeny from different regions of the United States. Individual samples were tested by hemagglutination-inhibition (HI) against six infectious bronchitis virus (IBV) strains: Massachusetts 41 (Mass), H52, Connecticut 46 (Conn), Arkansas 99, SE17, and JMK. The use of multiple strains to test broiler flocks resulted in the detection of seroconversions to Conn and JMK vaccination that were not detected with the IBV Mass HI test. Further, HI titers were detected to IBV strains not used for flock vaccination. In some cases, those titers could be due to cross reactions to antigens common to each of the virus strains. In two breeder flocks, the highest HI titers were to heterologous strains.     

Prevalence of Arkansas-type infectious bronchitis virus in Delmarva peninsula chickens

The prevalence of Arkansas (Ark)-type infectious bronchitis virus (IBV) in Delmarva peninsula broiler-type chickens was determined. The immunity of 5-to-11-week-old commercial broilers was evaluated by intraocular inoculation with Ark-type DPI strain (Ark DPI) challenge virus and collection of tracheal swabbings 5 days later. Serum Ark-type antibody titers were obtained using the virus-neutralization test. Eighty-five flocks were tested from January to August 1981. Nearly 60% of the flocks had substantial (greater than or equal to 70%) local immunity of the upper respiratory tract. Twenty-two percent had intermediate (50-69%) and 19% of the flocks had low (less than or equal to 40%) levels of local immunity. Serum antibody titers generally agreed with challenge results. In addition, high Ark-type IBV neutralizing-antibody titers were found in 16 Delmarva broiler breeder flocks. Seven current IBV field isolates were characterized for antigenic similarity to Ark DPI. Four isolates contained Ark antigen(s) based on significant neutralization in virus-neutralization tests and on substantial immunity to challenge afforded by Ark DPI virus immunization. Three isolates did not appear to contain Ark antigen(s). Immunization of chickens with Ark DPI virus afforded substantial protection against Connecticut- and homologous-type virus challenge, partial immunity (63%) against JMK, and no protection against the Massachusetts 41 strain of IBV.     

Detection of antibodies against serotypes 1 and 2 infectious bursal disease virus by commercial ELISA kits

Two distinct serotypes of infectious bursal disease virus (IBDV) are recognized in chicken and turkey flocks in the United States. Serologic testing of chicken flocks for serotype 1 viruses is routinely performed to monitor disease status and vaccination. Earlier studies indicated that enzyme-linked immunosorbent assay (ELISA) test detects antibodies to both serotypes of the virus, while the virus neutralization (VN) test is serotype specific. It is useful to evaluate currently available commercial ELISA kits for their ability to differentiate between antibodies elicited by the two serotypes. Three trials were performed in which chickens were orally inoculated with either a high or a low dose of serotype 1 STC or serotype 2 OH strains of IBDV. Sera collected at 0, 7, 14, and 21 days from these chickens and antisera procured from naturally infected broiler (n=20) and layer (n=30) flocks were tested with five different commercial ELISA kits and by VN. All ELISA kits detected different levels of antibodies elicited against serotype 1 of the virus and moderate and high levels of antibodies against serotype 2 virus. A correlation existed between the ELISA and the VN titers of experimentally infected chickens. All serum samples tested from the commercial layer flocks and 65% of the broiler flocks had antibodies against the OH strain. However, no correlation between the VN titers and ELISA titers was observed for the commercial broilers and layers sera by the majority of the kits. The results indicated that currently available commercial ELISA kits detect antibodies elicited by the two serotypes of IBDV. Hence, the prevalence of serotype 2 antibodies in the flocks should be considered while determining antibody profiles of the flocks against serotype 1 viruses.     

Serological profiles of commercial broiler breeders and their progeny. 2. Newcastle disease virus

Newcastle disease virus (NDV) hemagglutination-inhibition (HI) titers were determined for serum samples from eight commercial broiler breeder flocks and their progeny. The chickens sampled had been vaccinated and reared by different producers in different regions of the United States. Breeder flocks had the highest number of NDV-positive HI titers (greater than or equal to 1:10). Eighty percent or more of the samples from six of eight breeder flocks were positive; the geometric mean titers (GMTs) for those six breeder flocks ranged from 19 to 92. Only 3 of 8 broiler flocks had an increased frequency of positive titers and higher GMTs after vaccination. The frequency of positive titers was greater than 80% in only 2 of 8 of the oldest broiler flocks. The number of NDV-negative titers (less than 1:10) increased with age in most broiler flocks, even though all had been vaccinated once or more with live NDV vaccines.     

Infectious bronchitis serology in broilers and broiler breeders: correlations between antibody titers and performance in vaccinated flocks

In this study, a follow-up was made between 1993 and 1997 from broiler breeders at birth down to offspring broilers at processing, through vertically integrated registration of infectious bronchitis virus (IBV) antibody titers and performance data. All measurements were used two by two in a simple correlation study to calculate the degree to which they were linearly correlated. The antibody patterns in the broiler breeders indicated frequent field infections breaking through vaccinal immunity. Significant correlations measured between antibody titers and production parameters within and between the generations strongly suggested negative effects of IBV infections on laying percentage in the breeders and on mortality and daily weight gain in the broilers. Economic losses associated with IBV infections in the broilers occurred predominantly in flocks hatched with low and erratic maternal antibody titers. We concluded that IBV vaccination strategies should aim at high and uniform antibody titers in the broiler breeders.     

Inclusion body hepatitis as a primary disease in broilers in Saskatchewan, Canada

In recent years inclusion body hepatitis (IBH) has emerged as an economically important disease in Western Canada. Historically, infections with infectious bursal disease virus (IBDV) and chicken anemia virus (CAV) have been known to suppress the immune system of broilers and make them more susceptible to a secondary disease such as IBH. Recently it has been reported that virulent adenoviruses are able to cause IBH as a primary disease in broilers without apparent involvement of IBDV and CAV. The objectives of this study were to examine the possible association of IBH with IBDV and CAV infections in Western Canada and to identify adenoviruses involved in outbreaks. Serum samples from 17 broiler-breeder flocks and their progeny were collected when broilers were hatched and then again from broilers at the time of slaughter, and these samples were tested for IBDV and CAV antibodies by enzyme-linked immunosorbent assay (ELISA). Based on the ELISA titers the antibody response to vaccination against IBDV and CAV was at an expected level in all broiler flocks. Therefore, IBH outbreaks in these flocks were not due to inadequate levels of antibodies against IBDV and CAV. Moreover, there was no correlation found between occurrences of IBH outbreaks in broilers and their IBDV or CAV titers at the time of processing. Viruses that were isolated from livers of birds suffering from IBH could be classified into four different genotypes. Their hexon gene loop 1 sequences showed high percentages of identity to FAdV-7, FAdV-8a, FAdV-8b, and FAdV-11. The results of this study could not demonstrate an association of IBH with IBDV and CAV infections, but they supported the hypothesis that IBH in broilers in Western Canada is a primary disease with no apparent immunosuppressive involvement.     

Immunization of broiler breeder chickens against Newcastle disease with an oil-emulsion vaccine

Antibody response was rapid and high in broiler breeder chickens receiving 1 or 2 vaccinations with oil-emulsion vaccine against Newcastle disease at 23 or at 23 and 26 weeks old. The antibody titers remained high during the 41-week experimental period. At 64 weeks old, about 41 weeks after vaccination, the geometric mean hemagglutination-inhibition antibody titer was 67 from the single vaccination, and 103 from the double vaccination. The immune response to live-virus vaccine given at 2, 9, 20, 30, 42, or 54 weeks of age via the drinking water was high, but uniformity was lacking in the antibody response in the breeders and maternal antibody response in the progeny. Maternal antibody levels in one-day-old chicks were related to the titers of antibody in the dams. Maternal antibody titers of chicks originated from breeder flocks that were vaccinated with the oil-emulsion vaccine remained high for all hatches.     

Seroprevalence of avian influenza virus, infectious bronchitis virus, reovirus, avian pneumovirus, infectious laryngotracheitis virus, and avian leukosis virus in Nigerian poultry

Eight poultry farms in Nigeria, including chickens from nine breeder, 14 broiler, 28 pullet, 11 layer, and three cockerel flocks, were tested for antibody seroprevalence to the following poultry viruses of potential economic importance: infectious bronchitis virus (IBV), avian reovirus, avian pneumovirus (APV), infectious laryngotracheitis virus (ILTV), avian influenza virus (AIV), and avian leukosis virus (ALV). Serum samples were collected between 1999 and 2004 and were tested for antibodies using commercial enzyme-linked immunosorbent assay (ELISA) kits. Seroprevalence was very high for IBV (84%); intermediate for reovirus (41%), APV (40%), and ILTV (20%); and very low for ALV (<5%) antibodies. By commercial ELISA, the seroprevalence of antibodies against AIV was, in some flocks, up to 63%. However, more specific assays did not confirm AIV antibodies, indicating that all flocks tested were free of avian influenza antibodies. Birds seemed to be first infected by IBV (at about 7 wk of age), then by reovirus at 12 wk, before they became infected by APV (week 25) and ILTV (week 30). This is the first report of serological evidence of the above viruses in West Africa. Further studies are necessary to assess economic losses due to these avian viruses and the costs and benefits of countermeasures.     

Detection of chicken anemia virus in the gonads and in the progeny of broiler breeder hens with high neutralizing antibody titers

Previous evidence for the presence of chicken anemia virus (CAV) in the gonads of immune specific-pathogen-free chickens raised the question whether this occurs also in commercial breeders. The presence of CAV was investigated by nested PCR in the gonads and spleens of hens from two 55- and 59-week-old, CAV-vaccinated (flocks 2 and 3), and two 48- and 31-week-old non-vaccinated broiler breeder flocks (flocks 1 and 4). In addition, lymphoid tissues of 20-day-old embryos from these hens were also investigated for the presence of CAV. CAV was detected in the gonads and of 5/6 and 11/22 of the vaccinated hens and in some hens also in the spleen alone. Embryos from 7/8 and 5/18 of these hens were positive. In the non-vaccinated flocks, CAV was detected in the gonads of 11/34 and 10/10 hens in flocks 1 and 4, respectively. In addition, 11 birds in flock 1 had positive spleens. CAV DNA was detected in 3/11 and 2/10 of their embryos. CAV-positive gonads and embryos were detected in samples from hens with moderate as well as high VN antibody titers. Vaccinated chickens positive for CAV in the gonads and in their embryos had VN titers ranging from >1:512 to <1:2048. In non-vaccinated chickens, the VN titers of CAV positive chickens ranged from 1:128 to 1:4096. These results demonstrate that CAV genome can remain present in the gonads of hens in commercial broiler breeder flocks even in the presence of high neutralizing antibody titers that have been associated with protection against CAV vertical transmission. It also suggests that transmission to the progeny may occur irrespectively of the level of the humoral immune response in the hens.     

Comparison of a putative second serotype of chicken infectious anemia virus with a prototypical isolate I. Pathogenesis

CIAV-7 is a virus with similar pathogenic and physicochemical characteristics to, but antigenically distinct from, chicken infectious anemia virus (CIAV). The pathogenesis of CIAV-7 was evaluated in a comparative study with a representative isolate of CIAV, the Del-Ros strain. The pathogenesis of CIAV-7 was similar to Del-Ros on the basis of the clinical disease induced and gross and microscopic lesions, although CIAV-7 produced fewer and less severe lesions overall. A second comparative pathogenesis study was performed with Del-Ros and CIAV-7, both alone and in combination with infectious bursal disease virus (IBDV). In this study, the pathogenesis of CIAV-7 was similar to Del-Ros in clinical, gross, and microscopic lesions in the bone marrow. However, thymic lesions were less severe in CIAV-7-inoculated birds. The interaction between Del-Ros and IBDV was synergistic, whereas there was no observed potentiation of CIAV-7-induced disease by IBDV. Progeny from breeder flocks from several geographic locations in the eastern United States were challenged with CIAV-7 or Del-Ros to assess protection by maternal antibodies. Some progeny from all flocks had protection against CIAV-7 challenge, providing evidence for the presence of CIAV-7 in the field. Additionally, the number of birds protected against CIAV-7 or Del-Ros challenge varied within flocks, demonstrating that the agents are serologically distinct.     

A study of the epidemiology of infectious bursal disease in poultry in Queensland, 1976–1979

The epidemiology of infectious bursal disease (IBD) was studied by serology and sometimes by visual examination of the bursa of Fabricius in poultry flocks in Queensland during 1976–1979.
Ten flocks, each of approximately 30,000 meat breeding chickens, were surveyed. All chickens had maternally-derived antibody against IBD virus (IBDV) at hatching and active antibody was not detected while the chickens were brooded on rearing farms. When distributed to breeding farms, 7 of the flocks developed antibody when 11 to 25 weeks of age. The remaining 3 flocks were vaccinated by infection of 10% of the birds and within 4 weeks more than 80% of the chickens had developed precipitating antibody to IBDV.
Blood samples of 20 to 30 broiler chickens were collected at slaughter (7 to 9 weeks of age) from each of 312 broiler flocks raised on 37 contract farms. While the samples from 21 flocks were without detectable antibody to IBDV, all serum samples for 263 flocks contained antibody. The ratio of bursal weight to bodyweight was significantly lower in birds from 144 flocks having antibody to IBDV than in birds from 10 flocks that were without detectable antibody. In sequential studies, IBDV antibody became demonstrable in 27 of 30 flocks when the chickens were one to 6 weeks of age and was accompanied by bursal atrophy.
Serological investigation of 4 flocks of layer breeding chickens on a multi-age farm at approximately monthly intervals resulted in antibody to IBDV being detected at every examination.
Serological tests and bursal examinations were carried out weekly in 2 flocks each of 4000 layer chickens between one and 20 weeks of age. Serum antibody developed in one flock at 4 weeks of age and in the other at 17 weeks of age. In both flocks, bursal atrophy occurred concurrently with the development of antibody.     

Subclinical infectious bursal disease in an integrated broiler production operation.

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.     

Significance of infectious bursal disease serology in an integrated quality control program under European epidemiologic conditions

In this study performed between 1993 and 1997, infectious bursal disease virus (IBDV) antibody titers and performance data were recorded in a vertically integrated monitoring scheme in order to make a follow-up from day-old parents down to the broilers at slaughter. All measured data were used two by two in a simple correlation study to calculate the degree to which they were linearly correlated. It appeared that high and/or uniform antibody titers in the parents were correlated with increased daily weight gain and decreased mortality and slaughterhouse condemnation in the broilers. Antibody titers and their CVs were negatively correlated in broiler parents and their offspring at day-old and even at slaughter. Results indicate that high and uniform antibody titers against IBDV in broiler parents are important for good performance of the broiler offspring, at least under the epidemiologic conditions of this study, which included the presence of very virulent IBDV strains in the field and the sole use of live intermediate vaccines in broilers as well as broiler parents.     

Prediction of optimal vaccination timing for infectious bursal disease based on chick weight

Growth rate in broiler birds has increased substantially in the last decade due to improvement in genetics, feed formulation, cleaner environment, and vaccine formulations. As a result, it has become necessary to review and revise prediction method for vaccination in chicks. This study was undertaken to determine the possible use of the rate of weight gain rather than age in predicting vaccination time. Two groups of 1-day-old broilers originating from old and young breeders, respectively, and with different levels of maternal antibodies against infectious bursal disease virus (IBDV) were used in this study. The chicks were divided into four groups and subjected to two feed regiments: groups A1 and B1 were fed broiler feed for normal growth rate, and groups A2 and B2 were fed breeder feed for slower growth rate. At 1, 4, 8, 12, 16, 22, 29, and 36 days of age, 22 chicks in each group were weighed, and blood samples were collected. Serum samples were tested for antibodies against IBDV by enzyme-linked immunosorbent assay (ELISA) and virus neutralization test. Maternal antibody decline curves for each group were plotted according to chick age and chick weight. Fast-growing birds in groups A1 and B1 showed a faster rate of antibody decline, whereas slow-growing birds in groups A2 and B2 had a slower rate of antibody decline. Based on the effect of weight gain on maternal antibody decline, a new way of predicting vaccination time for IBDV based on measuring maternal antibody titers at 4 days of age was proposed and tested. The predicted antibody decline was shown to correspond to the real ELISA titers measured in our experiments (R = 0.9889), whereas a lower correlation (R = 0.8355) was detected between real ELISA titers and the titers predicted by the current method using age-based Deventer formula.     

Serosurvey of five viruses in chickens on smallholdings in Bangladesh

A serologic survey was undertaken in chickens in smallholdings in Bangladesh for avian influenza A virus (AIV), egg drop syndrome '76 virus (EDS'76V), infectious bronchitis virus (IBV), Newcastle disease virus (NDV) and reovirus (RV) in three phases: January 2002-May 2003, September 2003-August 2004, and August 2005-March 2006. Four hundred thirty-six sera collected in the 2nd phase, 295 in the first phase, 755 in the 1st plus 2nd phases and 295 in the 1st phase were investigated for AIV, EDS'76V, IBV and RV, respectively, using enzyme linked immunosorbent assays. All 854 sera collected in the three phases were screened for NDV using hemagglutination inhibition test. In chickens 20% were seropositive to AIV, 3% to EDS'76V, 74% to IBV, 88% to NDV, and 47% to RV. The seroprevalence in flocks was 23% to AIV, 6% to EDS'76V, 79% to IBV, 89% to NDV and 56% to RV. Twenty-five percent chickens had > or = 10log(2)HI titers to NDV.