Article history: Received: 26-11-2016, Accepted: 07-02-2017, Published online: 17-03-2017
Corresponding author: V. S. Dhaygude
E-mail: email@example.comCitation: Dhaygude VS, Sawale GK, Chawak MM, Bulbule NR, Moregaonkar SD, Gavhane DS (2017) Molecular characterization of velogenic viscerotropic Ranikhet (Newcastle) disease virus from different outbreaks in desi chickens, Veterinary World, 10(3): 319-323.
Aim: Diagnosis of velogenic viscerotropic Ranikhet disease from six different flocks of desi chicken in and around Mumbai by gross and histopathological examination, isolation of virus and molecular methods.
Materials and Methods: A total of 25 carcasses (varying between 2 and 6 carcasses from each flock) of six different flocks of adult desi chicken were subjected to necropsy examination for diagnosis of the disease during the span of a year (2014-2015). After thorough gross examination, the tissue samples were collected and processed for virus isolation and histopathological examination. The 20% tissue homogenate was inoculated into 9-day-old specific pathogen free (SPF) embryonated eggs. Mean death time (MDT) of embryos after inoculation and intracerebral pathogenicity index (ICPI) were used to judge velogenic nature of the virus. Newcastle disease virus (NDV) was isolated from six cases and confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) targeting the partial fusion protein gene of the viral genome.
Results: A total of 25 carcasses (varying between 2 and 6 carcasses from each flock) of six different flocks of desi chicken were presented for postmortem examination to Department of Veterinary Pathology, Bombay Veterinary College, Parel, Mumbai during 2014-2015. The gross and histopathological examination revealed lesions suggestive of viscerotropic velogenic form of the Newcastle disease (ND). The 20% tissue homogenate was inoculated into 9-day-old embryonated eggs from SPF chicken. NDV was isolated from six cases and confirmed by RT-PCR targeting the partial fusion protein gene. MDT of all the isolates was <60 h which indicated velogenic nature of the virus. ICPI of the isolates ranged between the 1.63 and 1.78. In four out of six outbreaks concurrent moderate to heavy infection of Ascardii galli in one flock and Railetina spp. in three flocks was also noted. In this study, viscerotropic velogenic form of ND was confirmed in all six outbreaks by gross and microscopic examination, virus isolation and RT-PCR.
Conclusion: In this study, viscerotropic velogenic form of ND was confirmed in all six outbreaks by gross and microscopic examination, virus isolation and RT-PCR. Nowadays, vaccine strains Lasota, B1 and F strains are used widely in India to control the infection of NDV. However, virulent NDV strains are still isolated frequently in the birds under backyard and also in commercial venture which demonstrates that NDV remains an on-going threat to commercial as well as backyard poultry flocks.
Keywords: mean death time, Newcastle disease virus, reverse transcriptase polymerase chain reaction, specific pathogen free, velogenic.
1. Beard, C.W. and Hanson, R.P. (1984) Newcastle disease. In: Hofstad, M.S., Barnes, H.J., Calnek, B.W., Reid, W.M. and Yoder, H.W., editors. Disease of Poultry. 8th ed. Iowa state University Press, Ames, IA. p452-470, 452-467.
2. Aldous, E.W., Mynn, J.K., Banks, J. and Alexander, D.J. (2003) A molecular epidemiological study of avian paramyxovirus Type 1 (Newcastle disease virus) isolates by phylogenetic analysis of a partial nucleotide sequence of the fusion protein gene. Avian Pathol., 32: 239-258. [Crossref] [PubMed]
3. Kim, S.H., Nayak, S., Paldurai, A., Nayak, B., Samuel, A., Aplogan, G.L., Awoume, K.A., Webby, R.J., Ducatez, M.F., Collins, P.L. and Samal, S.K. (2012) Complete genome sequence of a novel Newcastle disease virus strain isolated from a chicken in west Africa. J. Virol., 86: 11394-11395. [Crossref] [PubMed] [PMC]
4. Cattoli, G., Susta, L., Terregino, C. and Brown, C. (2011) Newcastle disease: A review of field recognition and current methods of laboratory detection. J. Vet. Diagn. Invest., 23(4): 637-656. [Crossref] [PubMed]
5. Hamid, H., Campbell, R.S.T. and Parede, L. (1991) Studies of the pathology of velogenic Newcastle disease virus: Virus infection in immune and nonimmune birds. Avian Pathol., 20: 561-575. [Crossref] [PubMed]
6. Alexander, D.J. (2003) Newcastle disease and other avian Paramyxoviridae infections. In: Shaif, Y.M., Barnes, H.J., Glisson, J.R., Fadly, A.M., McDougald, L.R. and Swayne, D.E., editors. Disease of Poultry. 12th ed. Blackwell, Oxford, UK. p75-100. [PubMed]
7. Aris, H., Vera, W., Pakpahan, S. and Wijayanti, N. (2016) Molecular pathotyping of Newcastle disease virus from naturally infected chickens by RT-PCR and RFLP methods. Asian J. Anim. Sci., 10: 39-48. [Crossref]
8. Asma, A., Mohammad, S.D., Muhammad, H., Mujahid, H., Shahid, M. and Khalid, A. (2016) Isolation, identification and molecular characterization of highly pathogenic Newcastle disease virus from field outbreaks. Braz. Arch. Biol. Technol., 59: 1. [Crossref]
8. Dash, S., Chakravarty, A.K., Singh, A., Upadhyay, A., Singh, M. and Yousuf, S. (2016) Effect of heat stress on reproductive performances of dairy cattle and buffaloes: A review. Vet. World, 9(3): 235-244. [Crossref] [PubMed] [PMC]
9. Singh, K., Jindal, N., Gupta, S.L., Gupta, A. and Mittal, D. (2005) Detection of Newcastle disease virus genome from the field outbreaks in poultry by reverse transcription - Polymerase chain reaction. Int. J. Poult. Sci., 4: 472-475. [Crossref]
10. Kiernan, J.A. (1999) Histochemical and Hitochemical Methods. 3rd ed. Butterworth/Heinemann, Oxford. p109-113, 200-210.
11. OIE, World Organization for Animal Health, (2012) In: Manual of Diagnostic Tests and Vaccines for Terrestrial animals. 7th ed. Ch. 2.3.14. OIE, Paris, France. p556-573.
12. Wakamatsu, N., King, D.J., Seal, B.S., Samal, S.K. and Brown, C.C. (2006) The pathogenesis of Newcastle a comparison of selected Newcastle disease virus wild-type strains and their infectious clones. Virology, 353: 333-343. [Crossref] [PubMed]
13. Kant, A., Koch, G. and Roozelaar, D. (1997) Differentiation of virulent and non-virulent strains of Newcastle disease virus within 24 hours by polymerase chain reaction. Avian Pathol., 26: 837-849. [Crossref] [PubMed]14. Nanthakumar, T., Kataria, R.S., Tiwari, A.K., Butchaiah, G. and Kataria, J.M. (2000) Pathotyping of Newcastle disease viruses by RT-PCR and restriction enzyme analysis. Vet. Res. Commun., 24: 275-286. [Crossref] [PubMed]