The present study describes the development of DNA vaccines using the hemagglutinin-neuraminidase (HN) and fusion (F) genes from AF2240 Newcastle disease virus strain, namely pIRES/HN, pIRES/F and pIRES-F/HN. Transient expression analysis of the constructs in Vero cells revealed the successful expression of gene inserts in vitro. Moreover, in vivo experiments showed that single vaccination with the constructed plasmid DNA (pDNA) followed by a boost with inactivated vaccine induced a significant difference in enzyme-linked immunosorbent assay antibody levels (p < 0.05) elicited by either pIRES/F, pIRES/F+ pIRES/HN or pIRES-F/HN at one week after the booster in specific pathogen free chickens when compared with the inactivated vaccine alone. Taken together, these results indicated that recombinant pDNA could be used to increase the efficacy of the inactivated vaccine immunization procedure.
Animals ; Antibodies, Viral/blood ; Cercopithecus aethiops ; Chickens ; *HN Protein/genetics/immunology ; Immunogenicity, Vaccine/*immunology ; Newcastle Disease/immunology ; Newcastle disease virus/enzymology/*genetics/immunology ; Specific Pathogen-Free Organisms ; Vaccines, DNA/genetics/*immunology ; Vaccines, Inactivated/immunology ; Vero Cells ; *Viral Fusion Proteins/genetics/immunology ; Viral Vaccines/genetics/*immunology/*standards

The ability of a heat-inactivated whole virus from a highly virulent infectious bursal disease virus (hvIBDV) and VP2 protein from hvIBDV expressed in E. coli provided protection against a hvIBDV challenge in specificpathogen- free (SPF) chickens. Six out of seven chickens that were injected three times with crude VP2 protein developed significant antibody titer against IBDV. However, only four out of the seven chickens survived the hvIBDV challenge. Despite showing low antibody titer profiles, all chickens immunized with the heat-inactivated whole virus also survived the challenged with hvIBDV. However, all of these chickens had bursal atrophy and mild to moderate depletion of lymphocytes. Thus, antibodies raised against IBDV VP2 protein expressed in E. coli and denatured IBDV proteins induced some degree of protection against mortality but not against bursal damage following challenge with hvIBDV.
Animals ; Antibodies, Viral/blood ; Birnaviridae Infections/immunology/prevention & control/*veterinary/virology ; Chickens ; Enzyme-Linked Immunosorbent Assay/veterinary ; Escherichia coli/genetics ; Immunization/standards/*veterinary ; Infectious bursal disease virus/genetics/*immunology/pathogenicity ; Poultry Diseases/*immunology/prevention&control/virology ; Recombinant Proteins/genetics/*immunology ; Specific Pathogen-Free Organisms ; Vaccines, Attenuated/immunology/pharmacology ; Vaccines, Synthetic/immunology/pharmacology ; Viral Structural Proteins/biosynthesis/genetics/*immunology ; Viral Vaccines/*immunology/pharmacology

A Newcastle disease virus (NDV) isolate designated IBS002 was isolated from a commercial broiler farm in Malaysia. The virus was characterised as a virulent strain based on the multiple basic amino acid motif of the fusion (F) cleavage site 112RRRKGF117 and length of the C-terminus extension of the hemagglutinin-neuraminidase (HN) gene. Furthermore, IBS002 was classified as a velogenic NDV with mean death time (MDT) of 51.2 h and intracerebral pathogenicity index (ICPI) of 1.76. A genetic distance analysis based on the full-length F and HN genes showed that both velogenic viruses used in this study, genotype VII NDV isolate IBS002 and genotype VIII NDV isolate AF2240-I, had high genetic variations with genotype II LaSota vaccine. In this study, the protection efficacy of the recombinant genotype VII NDV inactivated vaccine was also evaluated when added to an existing commercial vaccination program against challenge with velogenic NDV IBS002 and NDV AF2240-I in commercial broilers. The results indicated that both LaSota and recombinant genotype VII vaccines offered full protection against challenge with AF2240-I. However, the LaSota vaccine only conferred partial protection against IBS002. In addition, significantly reduced viral shedding was observed in the recombinant genotype VII-vaccinated chickens compared to LaSota-vaccinated chickens.
Amino Acids, Basic ; Animals ; Chickens* ; Genetic Variation ; Genotype* ; Malaysia ; Newcastle disease virus* ; Newcastle Disease* ; Vaccination ; Vaccines* ; Virulence ; Virus Shedding

Fowl adenovirus (FAdV) is distributed worldwide and causes economic losses in the poultry industry. The objectives of this study were to determine the hexon and fiber gene changes in an attenuated FAdV isolate from Malaysia in specific pathogen-free chicken embryonated eggs (SPF CEE) and its infectivity in commercial broiler chickens. SPF CEE were inoculated with 0.1 mL FAdV inoculum via the chorioallantoic membrane (CAM) for 20 consecutive passages. The isolate at passage 20 (E20), with a virus titer of 10(8.7)TCID₅₀/mL (TCID₅₀, 50% tissue culture infective dose), was inoculated (0.5 mL) into one-day-old commercial broiler chicks either via oral or intraperitoneal routes. The study demonstrated that 100% embryonic mortality was recorded from E2 to E20 with a delayed pattern at E17 onwards. The lesions were confined to the liver and CAM. Substitutions of amino acids in the L1 loop of hexon at positions 49 and 66, and in the knob of fiber at positions 318 and 322 were recorded in the E20 isolate. The isolate belongs to serotype 8b and is non-pathogenic to broiler chickens, but it is able to induce a FAdV antibody titer. It appears that molecular changes in the L1 loop of hexon and the knob of fiber are markers for FAdV infectivity.
Adenoviridae ; Amino Acids ; Chickens ; Chorioallantoic Membrane ; Eggs ; Fowl adenovirus A ; Liver ; Malaysia ; Mortality ; Ovum ; Poultry ; Serogroup ; Specific Pathogen-Free Organisms ; Viral Load

Background: The commercially available Newcastle disease (ND) vaccines were developed based on Newcastle disease virus (NDV) isolates genetically divergent from field strains that can only prevent clinical disease, not shedding of virulent heterologous virus, highlighting the need to develop genotype-matched vaccines Objectives: This study examined the efficacy of the NDV genotype-matched vaccine, mIBS025 strain formulated in standard vaccine stabilizer, and in carboxymethyl sago starch-acid hydrogel (CMSS-AH) following vaccination via an eye drop (ED) and drinking water (DW). Methods: A challenge virus was prepared from a recent NDV isolated from ND vaccinated flock. Groups of specific-pathogen-free chickens were vaccinated with mIBS025 vaccine strain prepared in a standard vaccine stabilizer and CMSS-AH via ED and DW and then challenged with the UPM/NDV/IBS362/2016 strain. Results: Chickens vaccinated with CMSS-AH mIBS025 ED (group 2) developed the earliest and highest Hemagglutination Inhibition (HI) NDV antibody titer (8log 2 ) followed by standard mIBS025 ED (group 3) (7log 2 ) both conferred complete protection and drastically reduced virus shedding. By contrast, chickens vaccinated with standard mIBS025 DW (group 5) and CMSS-AH mIBS025 DW (group 4) developed low HI NDV antibody titers of 4log 2and 3log 2 , respectively, which correspondingly conferred only 50% and 60% protection and continuously shed the virulent virus via the oropharyngeal and cloacal routes until the end of the study at 14 dpc. Conclusions The efficacy of mIBS025 vaccines prepared in a standard vaccine stabilizer or CMSS-AH was affected by the vaccination routes. The groups vaccinated via ED had better protective immunity than those vaccinated via DW.

Objective: This study examined the immune protection of immune-complex (Vaccine A) and attenuated live (Vaccine B) IBD vaccines in specific-pathogen-free (SPF) chickens against a novel Malaysian variant IBD virus (vaIBDV) challenge. Methods: One-day-old (n =75) SPF chickens were divided randomly into the following three groups of 25 chicks each: Control, Vaccine A, and Vaccine B groups. The vaIBDV strain, UPM1432/2019, was used for the challenge at 21 and 28days post-vaccination (dpv). Five birds from unchallenged and challenged groups were sacrificed seven days post-challenge, and blood, bursa, spleen, and cloacal swabs were collected. The IBD antibodies (Abs), lymphoid lesions, and viral load were determined. Results: The UPM1432/2019 virus induced bursal damage in vaccinated SPF chickens despite Ab titers. The mean Ab titers of the Vaccine A challenged group were significantly lower (p < 0.002) than in the unchallenged group at 28 dpv. The bursal indices of the vaccinated unchallenged groups did not differ significantly from those of the vaccinated challenged groups (p = 0.94). Microscopically, the bursae of the challenged groups showed significant atrophy. The bursal lesion score was higher (p < 0.05) in the control and Vaccine B challenged groups than the Vaccine A challenged group. The challenged group had a higher viral load than the vaccinated groups (p < 0.001). Conclusions and Relevance: Neither vaccine fully protected against a vaIBDV challenge, highlighting the limitations of current vaccines and the need for further research.

Objective: This study examined the immune protection of immune-complex (Vaccine A) and attenuated live (Vaccine B) IBD vaccines in specific-pathogen-free (SPF) chickens against a novel Malaysian variant IBD virus (vaIBDV) challenge. Methods: One-day-old (n =75) SPF chickens were divided randomly into the following three groups of 25 chicks each: Control, Vaccine A, and Vaccine B groups. The vaIBDV strain, UPM1432/2019, was used for the challenge at 21 and 28days post-vaccination (dpv). Five birds from unchallenged and challenged groups were sacrificed seven days post-challenge, and blood, bursa, spleen, and cloacal swabs were collected. The IBD antibodies (Abs), lymphoid lesions, and viral load were determined. Results: The UPM1432/2019 virus induced bursal damage in vaccinated SPF chickens despite Ab titers. The mean Ab titers of the Vaccine A challenged group were significantly lower (p < 0.002) than in the unchallenged group at 28 dpv. The bursal indices of the vaccinated unchallenged groups did not differ significantly from those of the vaccinated challenged groups (p = 0.94). Microscopically, the bursae of the challenged groups showed significant atrophy. The bursal lesion score was higher (p < 0.05) in the control and Vaccine B challenged groups than the Vaccine A challenged group. The challenged group had a higher viral load than the vaccinated groups (p < 0.001). Conclusions and Relevance: Neither vaccine fully protected against a vaIBDV challenge, highlighting the limitations of current vaccines and the need for further research.

Objective: This study examined the immune protection of immune-complex (Vaccine A) and attenuated live (Vaccine B) IBD vaccines in specific-pathogen-free (SPF) chickens against a novel Malaysian variant IBD virus (vaIBDV) challenge. Methods: One-day-old (n =75) SPF chickens were divided randomly into the following three groups of 25 chicks each: Control, Vaccine A, and Vaccine B groups. The vaIBDV strain, UPM1432/2019, was used for the challenge at 21 and 28days post-vaccination (dpv). Five birds from unchallenged and challenged groups were sacrificed seven days post-challenge, and blood, bursa, spleen, and cloacal swabs were collected. The IBD antibodies (Abs), lymphoid lesions, and viral load were determined. Results: The UPM1432/2019 virus induced bursal damage in vaccinated SPF chickens despite Ab titers. The mean Ab titers of the Vaccine A challenged group were significantly lower (p < 0.002) than in the unchallenged group at 28 dpv. The bursal indices of the vaccinated unchallenged groups did not differ significantly from those of the vaccinated challenged groups (p = 0.94). Microscopically, the bursae of the challenged groups showed significant atrophy. The bursal lesion score was higher (p < 0.05) in the control and Vaccine B challenged groups than the Vaccine A challenged group. The challenged group had a higher viral load than the vaccinated groups (p < 0.001). Conclusions and Relevance: Neither vaccine fully protected against a vaIBDV challenge, highlighting the limitations of current vaccines and the need for further research.

Objective: This study examined the immune protection of immune-complex (Vaccine A) and attenuated live (Vaccine B) IBD vaccines in specific-pathogen-free (SPF) chickens against a novel Malaysian variant IBD virus (vaIBDV) challenge. Methods: One-day-old (n =75) SPF chickens were divided randomly into the following three groups of 25 chicks each: Control, Vaccine A, and Vaccine B groups. The vaIBDV strain, UPM1432/2019, was used for the challenge at 21 and 28days post-vaccination (dpv). Five birds from unchallenged and challenged groups were sacrificed seven days post-challenge, and blood, bursa, spleen, and cloacal swabs were collected. The IBD antibodies (Abs), lymphoid lesions, and viral load were determined. Results: The UPM1432/2019 virus induced bursal damage in vaccinated SPF chickens despite Ab titers. The mean Ab titers of the Vaccine A challenged group were significantly lower (p < 0.002) than in the unchallenged group at 28 dpv. The bursal indices of the vaccinated unchallenged groups did not differ significantly from those of the vaccinated challenged groups (p = 0.94). Microscopically, the bursae of the challenged groups showed significant atrophy. The bursal lesion score was higher (p < 0.05) in the control and Vaccine B challenged groups than the Vaccine A challenged group. The challenged group had a higher viral load than the vaccinated groups (p < 0.001). Conclusions and Relevance: Neither vaccine fully protected against a vaIBDV challenge, highlighting the limitations of current vaccines and the need for further research.