Veterinary World

     Open access and peer reviewed journal  

ISSN (Online): 2231-0916

ISSN (Print): 0972-8988


Home l Editorial board l Instructions for authors l Reviewer guideline l Open access policy l Archives l FAQ

Open Access

Copyright: The authors. This article is an open access article licensed under the terms of the Creative Commons Attribution License

( which permits unrestricted use, distribution and reproduction in any medium, provided the work is properly cited.

Research (Published online: 21-03-2015)

20. Patho-epidemiological study on Genotype-XIII Newcastle disease virus infection in commercial vaccinated layer farms - J. H. Khorajiya, Sunanda Pandey, Priya D. Ghodasara, B. P. Joshi, K. S. Prajapati, D. J. Ghodasara and R. A. Mathakiya

Veterinary World, 8(3): 372-381



   doi: 10.14202/vetworld.2015.372-381


J. H. Khorajiya: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat - 388 001, India;

Sunanda Pandey: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat - 388 001, India;

Priya D. Ghodasara: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat - 388 001, India;

B. P. Joshi: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat - 388 001, India;

K. S. Prajapati: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat - 388 001, India;

D. J. Ghodasara: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat - 388 001, India;

R. A. Mathakiya: Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat - 388 001, India;


Received: 30-11-2014, Revised: 12-01-2015, Accepted: 16-01-2015, Published online: 21-03-2015


Corresponding author: Sunanda Pandey, e-mail:

Citation: Khorrajiya JH, Pandey S, Ghodasara PD, Joshi BP, Prajapati KS, Hodasara DJ, Mathakiya RA (2015) Patho-epidemiological study on genotype-XIII Newcastle disease virus infection in commercial vaccinated layer farms. Veterinary World 8(3):372-381.

Aim: The present research work was carried out to study the patho-epidemiological aspects of Genotype-XIII Newcastle disease virus (NDV) infection in commercial layer in and around Anand, Gujarat. As the outbreaks have reported in vaccinated flocks, it was felt necessary to study the disease with respect to its changing pathogenicity and relevant aspects.

Materials and Methods: The study comprised of patho-epidemiology of Newcastle disease (ND) by information collected from different layer farms suffering from the disease in relation to incidence pattern and mortality, duration of mortality, susceptible age, and loss due to production performance. Clinical signs were recorded based on observations. During postmortem, gross lesions were also recorded. For histopathological examination visceral organs according to lesions were collected in 10% formalin and processed slide stained by hematoxylin and eosin for microscopic examination. Cultivation of virus was done in embryonated specific pathogen-free (SPF) eggs of 9-11 days and isolation of virus was done for haemagglutination (HA) and haemagglutination inhibition (HI) test and to identify pathotype of virus by intracerebral pathogenicity index (ICPI) test to determine the virulence of virus. The Genotype-XIII NDV was confirmed by F gene sequence and whole genome sequence.

Results: During the study mortality due to ND was recorded in 13 layer flocks in spite of routine vaccination, which usually contain Genotype-II strain of virus. The mortality was observed as high as above 50% with an average of 21.21%. The susceptible age for disease was found to be 6-14 weeks. The duration of mortality observed was 23 days. The disease resulted in a significant reduction in body weight, feed intake and drop in egg production. Majority of the outbreaks appeared during extremely hot months of April to June. Greenish diarrhoea was frequently seen in birds that survived early in infection. Mortality continued for 2-3 weeks and reduced with appearance of torticollis. Gross lesions were characterized by multifocal to diffuse hemorrhages around proventricular glands, necrotic (diphtheritic) haemorrhagic ulcers throughout the intestine, disseminated multiple foci of necrosis and pin-point hemorrhages in the spleen parenchyma. The microscopic lesions include focal to diffuse hemorrhages, diffuse infiltration of mononuclear cells, necrosis, and degeneration in visceral organs. All the 13 farm samples (n=13) resulted in death of all the embryos following incubation up to 72 h post-inoculation. All the 13 allantois fluids from field samples along with F and R2B vaccine sample were found positive for HA activity, which was further confirmed by HI using known NDV serum. The values of ICPI were 2.0 which were indicative of velogenic nature of the field NDV strain.

Conclusion: The study indicated that presently available live and attenuated vaccines which include Genotype-II NDV have failed in protecting the flocks against Genotype-XIII and resulted in outbreaks with mortality above 50%. ICPI score of 2.0 confirmed that the present outbreaks were due to Genotype-XIII NDV, which is velogenic in nature.

Keywords: genotype, histopathology, intra-cerebral pathogenicity index, Newcastle disease.

1. The economic times (2013) access on 3rd march 2014.
2. Aldous, E.W. and Alexander, D.J. (2001) Detection and differentiation of new castle disease virus (Avian paramyxotype I). Avian. Pathol., 30: 117-128.
3. Nanthakumar, T., Kataria, R.S., Tiwari, A. K., Butchiah, G. and Kataria, J. M. (2000) Pathotyping of New castle disease virus by RT-PCR and restriction enzyme analysis. Vet. Res. Communication, 24: 275-286.
4. Edwards, J.J. (1928) A new fowl disease. Ann. Rep. Imp. Inst. Vet. Res. Mukhteswar 1978.
5. Kylasam, Aier. (1930) A study on Madras fowl pest. Ind. Vet. J., 8: 346-352.
6. International Committee on Taxonomy of Viruses, 9th Report. (2009), Virus Taxonomy: 2102 Release.
7. Briand, F.X., Henry, A., Masinand, P. and Jestin, V. (2012) Complete genome sequence of a novel avian paramyxovirus. J. Virol., 86(14):7710.
8. Susta, L., Jones, M. E. B., Cattoli., Cardenas-Garcia., Miller, P. J., Brown, C. C. and Afonso, C. L. (2014) Pathologic Characterization of Genotypes XIV and XVII Newcastle Disease Viruses and Efficacy of Classical Vaccination on Specific Pathogen-Free Birds. Vet. Pathol.,1: 2.
9. Luna, L. G. (1968) Manual of histologic staining methods of the armed forced institute of Pathology, 3rd edition New York, McGraw Hill Book Company.
10. OIE Terrestrial Manual. (2012) Newcastle disease. In: Manual of standards for diagnostic tests and vaccines.
11. Alexander, D.J. and Senne, D.A. (2008) Newcastle Disease and other Avian Paramyxoviruses. In: A Laboratory Manual for the Isolation, Identification and Characterization of Avian Pathogens, Dufour-Zavala L. (Editor in Chief) Swayne, D. E., Glisson, J.R., Jackwood, M.W., Pearson, J.E., Reed, W.M, Woolcock, P.R., 4th ed., American Association of Avian Pathologists, Athens, pp. 135-141.
12. Saidu, L. and Abdu, P.A. (2008) Outbreak of Viscerotropic Velogenic form of Newcastle dis-ease in vaccinated six weeks old pullets. Sokoto Journal of Veterinary Sciences, 7(1): 2
13. Gowthman, V., Singh, S. D., Dhama, K., Barathidasan, R., Srinivasan, P., Saravanan, S., Sukumar, K., Pandey, A.B. and Ramakrishnan, M.A. (2013) Pathology of Co-infections of low pathogenic avian influenza and virulent Newcastle disease in growers and layers-A field Observations. Compendium Abstracts Veterinary Pathology Congress. 3: 159.
14. Beach, J. R. (1943) Avian Pneumoencephalitis. Proc. 46th An. Meet., U. S. Livestock Sanitary Assoc, pp. 203-223.
15. Premavathi, K. and Vardhani, V.V. (2007) Incidence of Ranikhet disease in Broilers of organized farms in India. Ecology, Environment and Conservation Paper, 13(1): 187-190.
16. Njagi, L. W., Nyaga, P. N. and Uema, M. (2010) A retrospective study of factors associated with Newcastle disease outbreaks in village indigenous chickens. Bull. Anim. Health. Prod. Afr., 58: 22-33.
17. Leow, B. L., Shajarutulwardah, M. Y. and Ramlan, M. (2011) Newcastle disease in Malaysia: diagnostic cases in veterinary research institute (VRI) IPOH from 2004-2009. Malaysian Journal of Veterinary Research. 2(1) : 45-51
18. Tirumurugaan, K. G., Vinupriya, M. K., Vijayarani, K. and Kumanan, K. (2011b) Analysis of the fusion protein cleavage site of Newcastle disease virus isolates from India reveals preliminary evidence for the existence of II, VI and VII genotypes. Indian J. Virol., 22(2): 131–137.
PMid:23637515 PMCid:PMC3550736
19. Akamura, K. N., Htsu, N. O., Akamura, T. N., Amamoto, Y. Y., Amada, M. Y., Mase, M. and Imai, K. (2008) Pathologic and Immunohistochemical Studies of Newcastle Disease (ND) in Broiler Chickens Vaccinated with ND: Severe Nonpurulent Encephalitis and Necrotizing Pancreatitis. Vet. Pathology online.,45: 928–933.
20. Merino, R., Hilda, V., Jose, A., Quintana and Norma, C. (2011) Comparison of the Virulence of Pathogenic Newcastle Disease Viruses Belonging to the Same or Different Genotypes. Int. J. Poultry Sc., 10(9): 713-720.
21. Pazhanivel, N., Vijayalingam, T. A., Thangathurai, R., Kumar, V., Nithya, P. and Thanaseelan, V. (2013). Concurrent Newcastle disease and taeniasis in a coloured broiler chicken. Compendium Abstracts Veterinary Pathology Congress., 3: 153.
22. Wang, Yuyang., Zhiqiang, Duan., Shunlin, H., Yan, Kai., Xiaobo, Wang., Qingqing, Song., Lei, Zhong., Qing, Sun., Xiaoquan, Wang., Yantao, Wu. andXiufan, Liu. (2012) Lack of detection of host associated differences in Newcastle disease viruses of genotype VIId isolated from chickens and geese. Virol. J., 9: 197.
PMid:22971647 PMCid:PMC3491030
23. Hu, Zenglei., Hu, Jiao., Hu, Shunlin., Liu, Xiaowen., Wang, Xiaoquan., Zhu, Jie. and Liu, Xiufan. (2012) Strong innate immune response and cell death in chicken splenocytes infected with genotype VIId Newcastle disease virus. Virol. J., 9: 208.
PMid:22988907 PMCid:PMC3489799
24. Ezema, W.S., Okoye, J.O.A. andNwanta, J.A. (2009) LaSota vaccination may not protect against the lesions of velogenic Newcastle disease in chickens. Trop. Anim. Health Prod., 41(4): 477-84.
25. Tirumurugaan, K. G., Kapgate, S., Vinupriya, M. K., Vijayarani, K., Kumanan, K and Elankumaran, S. (2011a) Genotypic and pathotypic characterization of ND viruses from India. PLoS ONE 6(12): e28414.
PMid:22174801 PMCid:PMC3235129
26. Diel, D. G., Susta, L., Garcia, S. C., Killian, M. L., Brown, C. C., Miller, P. J. and Afonso, C. L. (2011) Complete Genome and Clinico-pathological characterization of a virulent Newcastle disease virus isolate from South America. J. Clin. Microbiol., 50(2): 378–387.
PMid:22135263 PMCid:PMC3264182
27. Fringe, R., Bosman, A. M., Ebersohn, K., Bisschop, S., Abolnik, C. and Venter, E. (2012) Molecular characterisation of Newcastle disease virus isolates from different geographical regions in Mozambique in 2005. Onderstepoort J. Vet. Res., 79(1), Art. #409, 7 pages.
28. Majed, H. M., Zahid, A. A. H., Kadhim, L. I., and Hasoon, M. F. (2013) Conventional and molecular detection of Newcastle disease and infectious Bursal disease in chickens. JWPR., 3(1): 05-12.
29. Haque, M. H., Hossain, M. T., Islam, M. T., Zinnah, M. A., Khan, M. S. R. and Islam, M. A. (2010) Isolation and detection of NDV from field outbreaks in broiler and layer chickens by RT-PCR. Bangl. J. Vet. Med., 8(2): 87–92.
30. Rakibul, H. A. K. M., Ali, M. H., Siddique, M. P., Rahman, M. M. and Islam, M. A. (2010) Clinical and laboratory diagnoses of Newcastle and Infectious bursal diseases of chickens. Bangl. J. Vet. Med., 8(2): 131-140.
31. Gowthaman, V., Singh, S. D., Dhama, K., Barathidasan, R., Anjaneya. And Ramakrishnan, M.A. (2011). Pathology and molecular diagnosis of Newcastle disease virus infection in broiler breeders, Indian J. Vet. Pathol., 35(2):168-170.
32. Ananth, R., Kirubaharan, J. J., Priyadarshini, M. L. M. and Albert, A. (2008) Isolation of Newcastle Disease Viruses of High Virulence in Unvaccinated Healthy Village Chickens in South India. Int. J. Poultry Sci., 7(4): 368-373.
33. Muhammad, M., Muhammad, A., Muhammad, T., Siamak, Z. and Mikael B. (2012) Genomic and biological characterization of a velogenic Newcastle disease virus isolated from a healthy backyard poultry flock in 2010. Virol. J., 9: 46.
PMid:22340092 PMCid:PMC3295720
34. Al-Habeeb, M. A., Mohamed, M. H. A. and Sharawi, S. (2013) Detection and characterization of Newcastle disease virus in clinical samples using real time RT-PCR and melting curve analysis based on matrix and fusion genes amplification. Vet. World, 6(5): 239-243.
35. Naveen, K. A., Singh, S. D., Kataria, M., Barathidasan, R. and Dhama, K. (2014) Molecular characterization and phylogenetic analysis of selected pigeon paramyxovirus type-l (PPMV-l) Indian isolates. J. Biol. Sci.,14(2): 134-141.
36. Kianizadeh, M., Ideris, A., Shahrabadi, M. S., Kargar, R., Pourbakhsh, S. A., Omar, A. R. and Yusoff, K. (1999) Biological and Molecular Characterization of Newcastle Disease Virus Isolated from Iran. Arch. Razi Ins. 50.