Development and optimization of virus neutralization test in chicken embryonated eggs for indirect identification of avian influenza and Newcastle disease virus

Avian viral problems have been consistently reported in commercial poultry of Pakistan causing heavy economic losses to the poultry farmers. Authentic idenfication and confirmation of the causative agent is always been question mark for the selection of vaccinal strain in this regard. Current study was therefore undertaken to optimize the virus neutralization test for the serological survey of vaccinated poultry particularly for avian influenza virus’s subtypes and Newcastle disease virus. Various physiochemical factors such as concentration of antigen and antibody, Incubation temperature and incubation period for in vitro and in-vivo reaction of antigen and antibody were optimized in chicken embryonated eggs. Serum samples were obtained from vaccinated breeder birds of five commercial poultry breeder companies and subjected for VNT using different concentration of three antigen and their respective homologous antibodies under optimized conditions. AIV H9 (EID50-1×109.0/ml) and NDV (EID50-1×108.2/ml) having biological titer of 10-7 /50ul HA units were neutralize with 10-2/50ul HIU of antibody and incubated at 37°C for 30 minutes was injected subsequently into 10 day old chicken embryo followed by incubation at 37°C for 38 hours showed ≥90% neutralizing specificity. Furthermore, sera obtained from five AIV-H9, AIV-H5 and NDV exposed commercial poultry farms revealed that Big bird broiler, Big bird breeders and A&S chicks are 100% sensitive and specific whereas, Gateway chicks and Waqas poultry breeders showed 100% homology for AIV-H5 virus but do not confers similarity with prevailing AIV-H9 and NDV field strains. Therefore, high sensitivity, reproducibility and specificity VNT, it could be a tool for indirect detection of homology between vaccinal strain and wild virus antigen using known antisera. Particularly, for those organisms possess natural ability to mutate in the adverse climatic conditions. Keywords: Virus neutralization test, Avian Influenza Virus, Newcastle Disease Virus, Sensitivity, Specificity

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Generation and Evaluation of Recombinant Thermostable Newcastle Disease Virus Expressing the HA of H9N2 Avian Influenza Virus

Viruses ◽

10.3390/v13081606 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1606

Author(s):

Xiaorong Zhang ◽

Zongyi Bo ◽

Chenchen Meng ◽

Yin Chen ◽

Chengcheng Zhang ◽

...

Keyword(s):

Influenza Virus ◽

Avian Influenza ◽

Newcastle Disease Virus ◽

Avian Influenza Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Transmembrane Domain ◽

Economic Losses ◽

Tail Domain ◽

H9n2 Avian Influenza Virus

H9N2 avian influenza virus (AIV) has become endemic in many countries, causing great economic losses when co-infected with other pathogens. So far, several live vaccines based on Newcastle disease virus (NDV) vectors expressing influenza hemagglutinin (HA) have been developed. However, the thermostable recombinant NDV is rarely reported. In this study, using a thermostable NDV rAHR09 strain as the vector, three recombinant NDVs expressing native HA, chimeric HA ectodomain with transmembrane domain/C-terminal cytoplasmic tail domain from fusion protein of NDV, and HA ectodomain were generated, designated rAHR09-HA, rAHR09-HAF, and rAHR09-HAE. The MDT value of three recombinant NDVs was above 120 h, their ICPI value was about 0.03, and the recombinant NDVs were still infectious when treated for 100 min under 56 °C, which demonstrated that the recombinant NDVs kept the lentogenic and thermostable nature of rAHR09. The immunization data showed that rAHR09-HA and rAHR09-HAF induced a higher HI antibody titer against H9N2 AIV and NDV. After being challenged with H9N2 AIV, the rAHR09-HA and rAHR09-HAF could significantly reduce the virus shedding in cloacal and tracheal swab samples. Our results suggest that rAHR09-HA and rAHR09-HAF might be vaccine candidates against H9N2 AIV.

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Prevalence of Newcastle Disease Virus and Avian Influenza Virus (H7N3) in Poultry at Karachi

RADS Journal of Biological Research & Applied Sciences ◽

10.37962/jbas.v11i1.260 ◽

2020 ◽

Vol 11 (1) ◽

pp. 1-6

Author(s):

Amjad Ali Channa ◽

Nazeer Hussain Kalhoro ◽

Zaheer Ahmed Nizamani ◽

Ayaz Hussain Mangi ◽

Jamila Soomro

Keyword(s):

Influenza Virus ◽

Avian Influenza ◽

Newcastle Disease Virus ◽

Avian Influenza Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Virus Isolation ◽

Economic Losses ◽

Rt Pcr ◽

Agar Gel

Background: Poultry is largest and rapidly growing sector of livestock in Pakistan. It is mainly influenced by viral pathogens such as Newcastle Disease Virus (NDV) and Avian Influenza Virus (H7N3). These viruses cause severe disease in poultry and leads to heavy economic losses throughout the world. The outbreaks of these pathogens have been increased in last few decades. Therefore, the study about antigenic prevalence is needed to know about the emergence of these pathogenic viruses, and to get rid of severe ailments associated with reduced poultry production. Objectives: To determine the prevalence of Newcastle Disease Virus (NDV), Avian Influenza Virus (H7N3) and co-infections in poultry flocks at Karachi. Methodology: For detection of NDV and H7N3, a total of 200 tracheal swabs were collected and tested through virus isolation (V.I); the sample with positive virus isolation were tested through agar gel precipitation (AGP) and then the RNA was isolated through TRI Reagent, which was further tested through reverse transcription polymerase chain reaction (RT-PCR). Results: The virus isolation showed that 58% of samples were positive for various viruses. Agar gel precipitation (AGP) revealed that the occurrence of NDV, H7N3 and ND+H7 were 50%, 8% and 38%, respectively. RT-PCR for F and HA gene of NDV and H7N3 confirmed the presence of NDV and H7N3 in the poultry. Conclusion: It is concluded that NDV and H7N3 are circulating in the flocks causing co-infections, therefore it is important to know the field challenge of viruses and to prepare vaccine of circulating serotype of virus to mitigate the rate of infection.

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SURVEILLANCE FOR NEWCASTLE DISEASE VIRUS, AVIAN INFLUENZA VIRUS AND MYCOPLASMA GALLISEPTICUM IN WILD BIRDS NEAR COMMERCIAL POULTRY FARMS SURROUNDED BY ATLANTIC RAINFOREST REMNANTS, SOUTHEASTERN BRAZIL

Brazilian Journal of Poultry Science ◽

10.1590/1806-9061-2015-0164 ◽

2016 ◽

Vol 18 (3) ◽

pp. 387-394 ◽

Cited By ~ 3

Author(s):

MB Guimarães ◽

R Hurtado ◽

CP Bello ◽

RET Vanstreels ◽

AJP Ferreira

Keyword(s):

Influenza Virus ◽

Avian Influenza ◽

Newcastle Disease Virus ◽

Avian Influenza Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Mycoplasma Gallisepticum ◽

Southeastern Brazil ◽

Poultry Farms ◽

Commercial Poultry

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Comparison of single 1-day-old chick vaccination using a Newcastle disease virus vector with a prime/boost vaccination scheme against a highly pathogenic avian influenza H5N1 challenge

Avian Pathology ◽

10.1080/03079457.2013.873111 ◽

2014 ◽

Vol 43 (1) ◽

pp. 68-77 ◽

Cited By ~ 7

Author(s):

Helena Lage Ferreira ◽

Fabienne Rauw ◽

Jean François Pirlot ◽

Frédéric Reynard ◽

Thierry van den Berg ◽

...

Keyword(s):

Avian Influenza ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Highly Pathogenic Avian Influenza ◽

Highly Pathogenic ◽

Pathogenic Avian Influenza ◽

Vaccination Scheme ◽

Avian Influenza H5n1 ◽

Influenza H5n1

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Protection and Differentiation of Infected from Vaccinated Animals by an Inactivated Recombinant Newcastle Disease Virus/Avian Influenza H5 Vaccine

Avian Diseases Digest ◽

10.1637/9166-876709-digest.1 ◽

2010 ◽

Vol 5 (s1) ◽

pp. e23-e24

Author(s):

Bernardo Lozano-Dubernard ◽

Ernesto Soto-Priante ◽

David Sarfati-Mizrahi ◽

Felipa Castro-Peralta ◽

Ricardo Flores-Castro ◽

...

Keyword(s):

Avian Influenza ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Recombinant Newcastle Disease Virus

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Interaction between avian influenza subtype H9N2 and Newcastle disease virus vaccine strain (LaSota) in chickens

BMC Veterinary Research ◽

10.1186/s12917-018-1689-4 ◽

2018 ◽

Vol 14 (1) ◽

Cited By ~ 3

Author(s):

Hany F. Ellakany ◽

Ahmed R. Gado ◽

Ahmed R. Elbestawy ◽

Hatem S. Abd El-Hamid ◽

Hafez M. Hafez ◽

...

Keyword(s):

Avian Influenza ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Vaccine Strain ◽

Virus Vaccine ◽

Newcastle Disease ◽

Newcastle Disease Virus Vaccine

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Evaluation of immunity and protection in chicken administered a developed vaccine against predominant Middle Eastern strains of genotypes VI and VII of velogenic Newcastle Disease Virus

10.21203/rs.3.rs-16483/v1 ◽

2020 ◽

Author(s):

Soonham Sami Yaghmoor` ◽

Taha Abdullah Kumosani ◽

Elie Kamil Barbour ◽

Othman Abubaker Baothman

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Humoral Immunity ◽

Newcastle Disease ◽

Middle Eastern ◽

Economic Losses ◽

Bivalent Vaccine ◽

Velogenic Newcastle Disease Virus ◽

Ifn Γ ◽

H Protein

Abstract Background The velogenic-Newcastle Disease Virus (v-NDV) causes an important disease in chicken, associated with serious economic losses to the global poultry industry. This research evaluated the immunity in broilers administered a developed bivalent vaccine, aiming at protection against predominant Middle Eastern strains of genotypes VI and VII of v-NDV. The completely randomized design implemented in this evaluation included eight treatments, differing in birds being administered or deprived of the developed vaccine, with a difference in type of challenge, either by v-NDV strain(s) of genotype VI, VII, or both. Vaccination was administered subcutaneously at 6 and 21 d of age, followed by an intra-pectoral challenge at the age of 28 d. Results The acquired humoral immunity by vaccinated and challenged birds to Hemagglutinin (H) protein was the highest at market age of 40 d, compared to challenged birds deprived of vaccination, and to vaccinates deprived of challenge (P<0.05). The same statistical difference pattern was obtained by the cell-mediated immunity (CMI), represented by birds’ level of serum IFN- γ . The type of challenge by either strain(s) of genotype VI, VII, or VI+VII did affect statistically the cross reactivity of acquired humoral immunity specific to H protein of homologous versus heterologous strains. The absence of humoral immunity and the low IFN- γ levels at 28 d of age in challenged birds deprived of vaccination lead to highest mortality, and lowest performance compared to vaccinates and challenged, vaccinates and deprived of challenge, and unvaccinated-unchallenged birds (P<0.05). Conclusions The developed bivalent vaccine was able to induce enough humoral and CMI responses, enabling protection of the broilers against production losses by each of the three types of v-NDV challenges. It is recommended to conduct future studies to evaluate such types of vaccines in chicken breeders and commercial layers, reared in various world’s zones with existing endemicity of v-NDV.

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