Live Infectious Bursal Disease Vaccine Research Articles

Efficacy of different live infectious bursal disease vaccines on antibodies to infectious bursal disease virus and Newcastle disease vaccine La Sota in ISA brown chicks

Aim: In this study, the efficacy of five commercial live infectious bursal disease (IBD) vaccines on antibody (Ab) decay and response to a very virulent IBD virus (vvIBDV) and Newcastle disease (ND) vaccine La Sota in ISA Brown chicks were assessed. Methods: Seven groups of chickens (A, B, C, D, E, F, and G) comprising 50 chicks each were used for the study. Groups A, B, C, D, and E were vaccinated with Bur-706, MB-Strain, B87-Strain, Bursa-B2K, and HIPRA live IBD vaccines, respectively, at 14 and 28 days of age (doa), and challenged with a vvIBDV at 35 doa. Groups F and G served as positive and negative controls, respectively. At 42 doa, all groups were vaccinated with ND vaccine La Sota. Blood was collected at 1, 7, 14, 21, 28, 35, 42, and 49 doa, and serum was harvested. The serum was analyzed for IBD virus (IBDV) Ab titres using enzyme-linked immunosorbent assay (ELISA) and ND Ab titres using a haemagglutination inhibition (HI) test. Results: Results revealed significantly (p < 0.05) higher ELISA IBDV Ab titres in groups A to E than in F from 21 to 49 doa, with the highest in group D. At 49 doa, there was significantly (p < 0.05) higher HI Ab titre in groups A to E than in F with the highest in group D. In conclusion, the live IBDV vaccines were immunogenic following vaccination. The vaccines decreased the decay of IBDV Ab and mitigated the decreased response to ND vaccine La Sota in the chicks following the vvIBDV challenge, with more effects induced by Bursa-B2K. Conclusion: The live vaccines (Bur-706, MB-Strain, B87-Strain, Bursa-B2K, and HIPRA) were immunogenic following vaccination.

Histopathological evaluation of the effects of live Infectious bursal disease vaccine originated from WF2512 strain on bursa Fabricius in the broilers

Infectious bursal disease (IBD) is a viral disease that causes significant economic losses in young chickens, characterized by lymphoid depletion and inflammation in the bursa Fabricius (BF). The incidence of the disease shows an increasing trend all over the world. Active and passive immunization is very important as well as strict hygiene measures in combating outbreaks. However, the fact that live-attenuated vaccines (mild, intermediate, hot) used for this purpose cause immunosuppression because of bursal damage is seen as an important limitation. In this study, it was aimed to histopathologically investigate the effects of commercial IBD vaccines originating from WF2512 (intermediate plus/hot, orally with drinking water) on BF under routine broiler rearing conditions. For this, BFs of 55 Ross 308 hybrid breed chickens (50 test, 5 controls) from five different broiler farms were used. In addition to standard vaccines, the IBD vaccine was given on day 15, and five samples from each farm were obtained 10 days later (25th day). After the first sampling, the second BF sampling was performed at the age of 38 days. Histopathological bursal lesion score was applied to evaluate the effectiveness of the vaccine. Accordingly, it was determined that the bursal lesion score, which increased slightly to moderately in the first samples, decreased in the second samples (27-61%). This was accepted as an indication that the bursal damage, which increased with IBD vaccine administration, diminishes over time and that histological regeneration was increased.

In ovo application of a live infectious bursal disease vaccine to commercial broilers confers proper immunity

ABSTRACT Infectious bursal disease (IBD) is an economically important disease of young chickens caused by the Avibirnavirus infectious bursal disease virus (IBDV). Besides biosecurity, vaccination is the most important measure for IBDV control. Sufficient levels of maternally derived antibodies (MDA) protect against early challenge and also interfere with the take of live conventional vaccines. Recently, the field surveys conducted in four countries, published by Ashash, U., Noach, C., Perelman, B., Costello, C., Sansalone, P., Brazil, T. & Raviv, Z. [(2019). In ovo and day of hatch application of a live infectious bursal disease virus vaccine to commercial broilers. Avian Diseases, 63, 713–720] using the MB-1 vaccine strain by in ovo application or sub-cutaneous route at the day of hatch seem to conflict with the rule that very early application of a conventional live vaccine in birds with significant levels of MDA has very little chance of a successful immune response. An in ovo vaccination-challenge controlled experiment with MB-1 vaccine was performed using commercial broilers with high levels of MDA against IBDV and a vvIBDV challenge at 22 or 36 days of age. Clinical signs, bursa-bodyweight ratios, histology, serology, RT–PCR, Sanger- and deep sequencing were used to study the efficacy and safety of the in ovo-applied MB1 vaccine in comparison to an established immuno-complex vaccine. The study findings confirmed that the in ovo application of the live MB-1 vaccine in commercial broilers was successful and induced full protection against a vvIBDV challenge at 22 and 36 days of age, demonstrated by the bursa lesion score and qPCR and IBDV genotyping. Comparable to the field studies, a delayed viral replication of 2–3 weeks, following the in ovo administration of the MB1 vaccine, was observed.

Comparison study between the efficacy of immune complex and conventionally live vaccine against Gumboro disease in broilers

This study aimed to evaluate the immune response and histological changes of two Gumboro disease vaccines. Two hundred, days old broilers were divided into 4 groups: group A was vaccinated with live attenuated infectious bursal disease (IBD) vaccine at 7 and 21 days of age, group B was vaccinated with Immune-Complex vaccine (Icx) at 1 day old, then all broilers of the groups A, B and C were vaccinated with ND vaccine at 10 and 24 days, while the group D was negative control. The blood was collected at 1, 7,14,21,28 and 35 days of age to obtain serum for ELISA. Samples of bursa from broilers of all groups at 14 and 28 days of age were submitted for histological examination. As a result of vaccination in group A the antibody titers are elevated after the 1st and 2nd dose of vaccination at 7 and 21 days. In group B this titer is increased from 21 day of age and reaches to peak at 35 day 7810±858 with significant difference, while in unvaccinated groups C and D the titer decreased gradually. The histological examination of bursal sections in group A and B varied at 14 and 28 days post vaccination and showed degeneration and necrosis of follicular lymphocytes compared with group and D. Thus we conclude that (Icx) vaccine improves the immune response after IBD and ND vaccination in comparison with live IBD vaccine.

Efficacy of a novel in ovo-attenuated live vaccine and recombinant vaccine against a very virulent infectious bursal disease virus in chickens.

Infectious bursal disease (IBD) causes severe economic damage to the poultry industry worldwide. To prevent IBD virus (IBDV) infection, live virus vaccines have been widely used in chickens having wide-ranging levels of maternally derived antibodies. But, the risks of infection with other pathogens because of lesions related to atrophy of the bursa of Fabricius in vaccinated chickens are a concern. To resolve the problems, a recombinant turkey herpesvirus (HVT) vaccine expressing IBDV-VP2 protein (rHVT-IBD) has been developed. However, the induction of neutralizing antibodies by rHVT-IBD against a virulent IBDV might be delayed compared with that by the live IBD vaccine, leading to the high risks of IBDV infection for young chickens. To find the best selection of IBDV vaccine for the onset of immunity, we examine the protective efficacy of a novel in ovo-attenuated live IBDV (IBD-CA) vaccine and the rHVT-IBD vaccine in young chickens challenged with a very virulent IBDV (vvIBDV) strain. We show that the protective efficacy of IBD-CA vaccine was higher than that of the rHVT-IBD vaccine in 14-day-old chickens challenged with the vvIBDV strain, leading to the risk of IBDV infection for young chickens when vaccinated with rHVT-IBD. Our results suggest that farmers should select the best vaccines to maximize vaccine efficacy in consideration of the vaccine characteristics, prevalence levels of IBDV in the areas, and initial MDA levels of the chickens since the attenuated live and recombinant vaccines play a role in the different vaccine efficacies.

Assessment of the role of intracloacal inoculation of live infectious bursal disease vaccine in breaking through maternally derived antibodies

ABSTRACT Infectious bursal disease (IBD) remains a potential worldwide threat to the poultry industry despite several vaccination approaches. Because maternally derived antibodies (MDA) constitute a critical problem for IBD vaccination, we examined the efficiency of the intracloacal vaccination approach in breaking through MDA. Experiment 1 determined the ability of the vaccinal strain to multiply in the bursa of Fabricius (BF) in chicks with a high level of MDA. Using real-time polymerase chain reaction, we quantified the strain in the bursae of vaccinated and non-vaccinated chicks. Experiment 2 was performed on three groups of chicks with high levels of MDA: group 1, non-vaccinated non-challenged; group 2, non-vaccinated challenged; and group 3, vaccinated challenged. Seroconversion to IBDV was measured using enzyme-linked immunosorbent assay. Groups 2 and 3 were challenged by vvIBDV at 25 days of age. Experiment 3 studied the effect of early IBD vaccinal strain multiplication on the immune response of vaccinated and non-vaccinated chicks to other vaccines. In experiment 1, the vaccinal strain showed progressive multiplication and reached the detectable titre in BF at 12 h post-vaccination despite high MDA titre. Experiment 2 showed that chicks in group 3 had significant seroconversion against IBDV. After challenge, group 3 showed significant improvements in several measured parameters compared with group 2. Moreover, results of experiment 3 proved that early multiplication of the vaccinal strain in the BF has no significant effect on the immune system or immune response to other vaccines. These results proved the promising success of this IBD vaccination approach. RESEARCH HIGHLIGHTS IBD vaccinal strain succeeded in multiplying in BF after intracloacal inoculation. Vaccinated chicks showed significant seroconversion of IBDV antibody titres. Vaccinated chicks showed a significant protection level against vvIBDV. Early IBD vaccination did not affect the immune response to other vaccines.

Comparative Evaluation of HVT-IBD Vector, Immune Complex, and Live IBD Vaccines against vvIBDV in Commercial Broiler Chickens with High Maternally Derived Antibodies.

Simple SummaryVaccination is the main method to control infectious bursal disease (IBD) in commercial broilers worldwide. The main obstacle to the vaccination process is maternally derived antibodies; thus, new generations of vaccines such as vector and immune complex vaccines have been developed. The efficacies of new and classical vaccines were compared to those of vvIBDV in the presence of high levels of maternally derived antibodies. The best results were obtained when using the vector IBD vaccine followed by the immune-complex vaccine, and the use of killed along with the live intermediate vaccine in terms of mortality, feed conversion ratio, bursal and spleen index, bursal lesion score, and serology.Infectious bursal disease (IBD) causes increased mortality and severe immunosuppression in commercial chickens. Currently, vaccination mainly used to control IBD. In this study, Group A (n = 30) received the HVT-IBD vector vaccine (Vaxxitek®) s/c and Group B (n = 30) received the immune complex vaccine (Bursa-Plex®) s/c at 1 day of age. Group C (n = 30) received a single dose of intermediate plus vaccine (228E) through the eye-drop route at 14 days of age. Group D (n = 30) was vaccinated twice with the intermediate vaccine (D78) at 12 and 22 days of age by eye-drop. Group E (n = 30) had the same treatment as group D along with the IBD killed vaccine (Nobilis G®) at 5 days of age. The PC (n = 20) and NC (n = 20) groups were non IBD vaccinated birds either challenged or not with vvIBDV, respectively; 20 chicks from each group were challenged with vvIBDV at 4 weeks of age. Based on clinical signs, postmortem gross lesions, histopathological changes, mortality rate, feed conversion rate, serology, bursal and spleen indices, the HVT-IBD vector vaccine administered was found to be safer and provided better protection against the vvIBDV challenge. The use of a killed IBD vaccine at an earlier age in broilers strengthened the protection induced by double doses of intermediate vaccines in broilers with high maternally derived antibodies against the vvIBDV challenge.

Stinging nettle and neem enhance antibody response to local killed and imported live infectious bursal disease vaccines in indigenous chicken in Kenya

Immune responses are critical for protection of chickens from infectious bursal disease (IBD). In this study, the antibody response-enhancing effect of drinking water supplementation of 1% stinging nettle and neem on different IBD vaccines and vaccination regimes was evaluated, using 36 (n = 36) specific antibody negative indigenous chicks. The birds were allocated into 3 groups as follows: 1A-C, 2A-C, and 3A-B, while group 3C acted as the unvaccinated non-supplemented control. A local inactivated K1 and imported live attenuated D78 IBD vaccines were given to groups 1A-C and 3A-B at 14 and 28 d of age, respectively. A combination of K1 and D78 vaccines was given 30 d apart to groups 2A and 2B (D78 at 14 and 21 d and K1 at 44 d of age) and on the same d to group 2C at 14 and 28 d of age. Stinging nettle was given in water to groups 1B, 2B, and 2C, and neem to groups 1C, 2A, and 3B. Birds were bled weekly and immune responses monitored using indirect ELISA. Both neem and stinging nettle had antibody response-enhancing effects in groups 1B and 1C, receiving the local inactivated K1 vaccine. There were significant differences (P < 0.05) in antibody titers between groups 1A and 2C. Stinging nettle induced earlier onset of high antibody responses in group 2C and persistent titers (>3.8 log10) from the third week in group 2B. Imported live D78 vaccine induced higher antibody titers compared to the local inactivated K1 vaccine. Groups 2B and 2C receiving a combination of the local K1 and imported live attenuated D78 vaccines had the highest antibody titers. Adoption of stinging nettle supplementation and a prime-boost program involving use of a local virus isolates-derived vaccine is recommended.

Effect of Age and Route of Administration on the Efficacy of Live Infectious Bursal Disease Vaccines in Broiler

Effect of Age and Route of Administration on the Efficacy of Live Infectious Bursal Disease Vaccines in Broiler

Evaluation of immune responses by live infectious bursal disease vaccines to avoid vaccination failures.

Infectious Bursal Disease (IBD) is a viral, contagious immunosuppressive disease posing an important threat to the commercial poultry industry. Evolution of highly virulent strains of IBD virus warranted the need for detailed characterization of the immune responses offered by the currently available vaccines. Two extensively used live vaccines of varied attenuation levels - intermediate and intermediate plus - strains were analyzed for the induction of immune responses. Both the vaccines induced protective antibody titers with the onset, quicker and higher with the intermediate plus vaccine. The intermediate plus strain vaccinate was observed to induce higher levels of IFN-γ in the birds. These results were supported by immunophenotype analyses with an increase in CD8+ and simultaneous decrease in CD4+ cell population. Both vaccine strains conferred protective immunity against virulent challenge. The study warrants the use of intermediate plus vaccines in disease endemic regions and intermediate vaccines in non-endemic regions to prevent IBD infection.

Effects of Live Infectious Bursal Disease Vaccines, on Immune Response of Vaccinated Chicks

Aim: Relationship between virus titers of live Infectious Bursal disease (IBD) vaccines and their serum-conversion abilities was studied. Study design and Methodology: Five batches of each, of five IBD vaccine brands used in Nigeria, were tested for virus titers. Each of the vaccine brands was also used to vaccinate a group of fifteen 12-days old chicks to study their serum-conversion abilities. Mean antibody titers of the groups of chicks were plotted, on a graph, against virus titers of the vaccine brands used to vaccinate them. Results: Mean Modified Passive Haemagglutination titers of IBD virus in the vaccines,were:1,065.60±780.03,1,472.00±748.55,2,112.00±1984.00,2,176.00±1920. 00 and 2,585.00±926.92 while mean antibody titers they elicited were, 1,356.80±241.51, 1,280.00±174.88, 448.00±79.25, 998.40±196.27 and 332.80±51.20, respectively. Line of best fit of graph of antibody titers of vaccinated chicks on vaccine titers, showed that reducing titers of the live IBD vaccines improved their immunogenicity. Conclusions: The inverse relationship between virus titers of the vaccines and their Original Research Article British Journal of Medicine & Medical Research, 4(7): 1506-1513, 2014 1507 serum conversion abilities, suggests that, if viral titers of live IBD vaccines are too high, immune-suppression instead of enhancement of immune response may occur.

Field efficacy trial of a novel HVT-IBD vector vaccine for 1-day-old broilers

Two vaccination programmes for infectious bursal disease (IBD) were compared in broiler chickens with maternal immunity, placed on two farms. A turkey herpes virus (HVT)-IBD vector vaccine was administered by the subcutaneous route, at the hatchery, into the chicks of farm A at the age of 1 day. On farm B, an attenuated intermediate live IBD vaccine was given orally at the ages of 17 and 24 days. The vaccine uptake was monitored via serology and bursa/body weight ratio evolution, as well as PCR-based viral IBDV detection in the bursa of Fabricius at various time points. It was also verified by an experimental very virulent IBDV challenge performed at the age of 30 days in birds transferred from the farms with appropriate control groups in a laboratory. An immunity gap was observed in birds from farm B between the decay of the passive and the rise of the active immunity based upon serological data. The level of protection against challenge is not possible to establish in this farm as the reduction of the bursa/body weight ratio observed could be due to the residual pathogenicity of the vaccine strain or the challenge as well. This immunity gap was not present on farm A showing higher serological titres at the ages of 26 and 45 days via a suitable ELISA test and 93% protection against the very virulent challenge at the age of 30 days was observed. The maternal immunity interfering with the live IBDV vaccine replication had no detectable effect on the vector vaccine take.

The efficacy of live infectious bursal disease vaccines in commercial 10 days old chicks.

The efficacy of live infectious bursal disease vaccines in commercial 10 days old chicks.

Extensive antigenic changes in an atypical isolate of very virulent infectious bursal disease virus and experimental clinical control of this virus with an antigenically classical live vaccine

The 99323 Egyptian isolate of infectious bursal disease (IBD) virus (IBDV) was identified during an international survey of acute IBD cases. Its unique antigenicity was characterized by a markedly reduced binding of neutralizing monoclonal antibodies 3, 4, 5, 6, 8 and 9 in an antigen-capture enzyme-linked immunosorbent assay. Nucleotide sequencing of the genome region encoding the VP2 major immunogenic domain in 99323 revealed amino acid changes occurring at positions critical for antigenicity, but phylogenetic analysis demonstrated that 99323 was related to typical, very virulent IBDV (e.g. isolate 89163). Protection experimentally afforded by an antigenically classical live IBD vaccine was investigated in specific pathogen free chickens challenged with 99323 or 89163. Both viruses were similarly controlled, as evaluated by clinical signs, growth retardation, bursa-to-body weight ratios and histological lesions of the bursa after challenge. These results document that an active antibody response to a classical live antigen may clinically control infection by an antigenically atypical very virulent IBDV.

Effects of early age feed restriction and heat conditioning on heat shock protein 70 expression, resistance to infectious bursal disease, and growth in male broiler chickens subjected to heat stress

The effects of early age feed restriction and heat conditioning on heat shock protein (HSP) 70 expression, antibody production, resistance to infectious bursal disease (IBD), and growth of heat-stressed male broiler chickens were investigated. Chicks were divided into 4 groups: 60% feed restriction on d 4,5, and 6 (FR); exposure to 36 +/− 1 degrees C for 1 h from d 1 to 21 (HT); combination of FR and HT (FRHT); and control. From d 35 to 50, heat stress was induced by exposing birds to 38 +/− 1 degrees C and 80% RH for 2 h/d. On d 36, each bird was administered 10 times the normal dose of live IBD vaccine. After heat exposure, the FRHT birds had higher HSP 70 density (d 41) and weight gain (from d 35 to 49) and lower bursal histological score (BHS) (d 51) than their HT and control counterparts. The HSP 70 expression and BHS of FR birds were not significantly different from those of the other 3 groups during the heat exposure period. Heat shock protein 70 and BHS data were negatively correlated (r = −0.33, P = 0.0008). We concluded that FRHT could improve weight gain and resistance to IBD in male broiler chickens under heat stress conditions. The improved heat tolerance and disease resistance in FRHT birds could be attributed to better HSP 70 response.

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)

Vaccination of commercial layer chicks against infectious bursal disease with maternally derived antibodies

Day-old commercial layer chicks with maternally derived antibody to infectious bursal disease (IBD) were reared in four pens on a commercial farm, and chicks from the same flock were reared at the laboratory and examined for the decay of the maternal antibody and the development of susceptibility to IBD. The chicks in the pens on the farm were vaccinated with different combinations of commercial inactivated and live vaccines against IBD, transferred to the laboratory, and challenged with virulent IBD virus to assess the efficacy of the vaccination programmes. The chicks from one pen, given a live IBD vaccine only, did not develop antibodies and were fully susceptible to challenge by seven weeks old; only one to two weeks later than the unvaccinated control chicks reared at the laboratory. The chicks in the other three pens were inoculated with an inactivated vaccine, and the chicks in two of these pens were inoculated later with a live vaccine. All these chicks developed IBD antibodies and were protected by between 91 and 100 per cent against challenge. There was no demonstrable benefit in administering a live vaccine either alone or in addition to an inactivated oil emulsion vaccine.

A Comparison of Three Different Regimens of Infectious Bursal Disease Vaccination in Chickens

Five days old progeny chicks from breeders which have received primary and booster doses of live infectious bursal disease vaccine, demonstrated precipitating antibodies unlike progenies from single dose vaccinated breeders. All chicks from the two different groups of breeders were however seronegative at 22 days of age, despite vaccination at 7 or 14 days of age. Post vaccination seroconversion was first noticeable at 32 days in the group of chicks vaccinated at 7 days. Post challenge mortalities was significantly lower (P less than 0.05) and organ lesions very significantly minimized (P less than 0.01) in 7 day old vaccinated group than in 14 or 28 days old vaccinated chicks. These results showed that early (7 days) IBD Vaccination was superior to vaccination at 14 or 28 days, in terms of antibody response and protectivity against mortalities and bursal lesions.

Evaluation of the Immunogenicity, Stability, Pathogenicity, and Immunodepressive Potential of Four Commercial Live Infectious Bursal Disease Vaccines

The study was divided into three experiments. In each, day-old specific pathogen free (SPF) White Leghorns were injected subcutaneously with one of the following commercial live infectious bursal disease (IBD) vaccines: Burcell®, Bursine®, Clone Vac D-78®, and S-706®. In the first experiment, the immunogenicity of each vaccine was determined by challenge with virulent IBDV at 25 days of age. All four vaccines were equally efficacicous in preventing clinical IBDV infection. The second experiment determined the stability and pathogenicity of the four vaccines by successive back passage of each virus in SPF chicks for 4 consecutive weeks. Although all vaccines were capable of spreading to contact controls, only the two intermediate vaccines (D-78® and S-706®) produced slightly atrophied bursae and moderate microscopic bursal lesions. However, both vaccines were stable, because neither reverted back to increase virulence nor resulted in morbidity or mortality associated with virulent IBDV after successive passage. In the third experiment, the immunodepressive potential of each vaccine virus was determined by examining the ability of IBD-vaccinated birds to respond to Newcastle disease (ND) virus vaccination. None of the four vaccines was found to be immunodepressive, as all IBD vaccinated birds responded to NDV vaccination.

Vaccination of Day-Old Broiler Chicks against Newcastle Disease and Infectious Bursal Disease Using Commercial Live and/or Inactivated Vaccines

Day-old broilers were administered live and/or inactivated vaccines to assess vaccine efficacy against challenge with Newcastle disease (ND) and infectious bursal disease (IBD). Chicks were from commercial breeder pullets vaccinated against ND and IBD using several live vaccine primers followed by an inactivated ND-IBD vaccine at 18 weeks. The most efficacious initial ND-IBD vaccination program was live ND virus by eye drop and live IBD vaccine injected subcutaneously (SQ) followed 2 hours later with inactivated ND-IBD vaccine SQ. The next two most efficacious programs were live vaccine alone and the inactivated vaccine only. Inactivated vaccine given SQ had no adverse effect on live IBD vaccine given 2 hours earlier in a similar site. Administration of inactivated vaccine by vent was not as efficacious as administration SQ. A booster of a second live ND-IBD vaccine drinking water at 18 days significantly increased levels of circulating antibody, regardless of the initial vaccination program.