Isolation, molecular detection and BHK-21 adaptation of Newcastle disease virus of field cases in layer farms of Bangladesh

Authors

  • Mohammad Ferdousur Rahman Khan Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh- 2202, Bangladesh
  • Sultan Ahmed Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh- 2202, Bangladesh
  • M. Liakot Hossen Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh- 2202, Bangladesh
  • M. A. Imran Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh- 2202, Bangladesh
  • K. H. M. Nazmul Hussain Nazir School of Basic Studies, Yeungnam University, Gyeongsan 712-749, Republic of Korea
  • Marzia Rahman Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh- 2202, Bangladesh
  • M. T. Rahman Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh- 2202, Bangladesh
  • M. B. Rahman Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh- 2202, Bangladesh

Keywords:

NDV; ‘F’ gene; BHK-21 cell line; RT-PCR; Nested PCR

Abstract

In this study, field samples (n=45) such as trachea, brain and proventriculus were collected from 12 layer farms of different districts of Bangladesh for the detection and isolation of pathogenic Newcastle disease virus (NDV). Isolation of the virus from the clinical samples and their propagation was primarily carried out in chicken embryo. Among the forty five samples, 9 (20%) were found positive for NDV by hemagglutination (HA) and hemagglutination inhibition (HI) tests. The positive isolates were cultured in chicken embryo, and subsequently confirmed by reverse transcription-polymerase chain reaction (RT-PCR) targeting NDV specific ‘F’ gene that encodes fusion protein. A nested PCR was carried out with the RT-PCR product as the template, which targeted a smaller internal region of ‘F’ gene. The nested PCR also confirmed all the isolates as NDV. For further confirmation, NDV specific ‘F’ gene was also amplified from the cDNA generated from the RNA extracted from the virus inoculated chicken embryo by direct PCR. The isolates were inoculated in BHK-21 cell line and 2/3 blind passage were given without any gross change in the cell. Later, the virus was found adapted into BHK-21 cell line where they produced syncitia, rounding of cell and multinucleated giant cells as the cytopathic effects. Adaptation of the virus in BHK-21 cell line was confirmed by RT-PCR and nested-PCR successfully.

References

Ahamed, T., Hossain, K.M., Billah, M.M., Islam, K.M.D.,

Ahasan, M.M., Islam, M.E., 2004. Adaptation of Newcastle disease virus (NDV) on vero cell line. Int. J. Poult. Sci., 3, 153-156.

Aldous, E.W., Mynn, J.K., Banks, J., 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.

Alexander, D.J., 1997. Newcastle disease and other avian paramyxoviridiae infections. BW Calnek, HJ Barnes, CW Beard, LR McDougald and YM Saif (eds); Diseases of poultry. 10th edition, Iowa State University Press, Ames, Iowa, USA: pp 553-554.

Amin, M.A., Amin, M.M., Khan, M.S.R., Choudhury, K.A., Siddiky, M.N.A., Sarker, A.J., 2004. Characterization of Newcastle disease virus isolates from caged birds in Bangladesh. Bangl. J. Vet. Med., 2, 113-116.

Anon, 1971. Methods for examining poultry biologics and for identifying and quantifying avian pathogens. In: Newcastle disease, National Academy of Science, Washington DC; p. 66.

Ballagi-Pordany, A., Wehmann, E., Herczeg, J., Belak, S., Lomniczi, B., 1996. Identification and grouping of Newcastle disease virus strains by restriction site analyses of a region from the ‘F’ gene. Arch. Virol., 141, 243-261.

Biswas, P.K., Biswas, D., Ahmed, S., Rahman, A., Debnath, N.C., 2005. A longitudinal study on the incidence of major endemic and epidemic diseases affecting semi-scavenging chickens reared under the Participatory Livestock Development Project areas in Bangladesh. Avian Pathol., 34, 303-312.

Collins, M.S., Bashiruddin, J.B., Alexander, D.J., 1993. Deduced amino acid sequences at the fusion protein cleavage site of Newcastle disease viruses showing variation in antigenicity and pathogenicity. Arch. Virol., 128, 363-370.

Creelan, J.L., Graham, D.A., McCullough, S.J., 2002. Detection and differentiation of pathogenicity of avian paramyxovirus serotype-1 from field cases using one-step reverse transcriptase – polymerase chain reaction. Avian Pathol., 31, 493-499.

Czermak, P., Pörtner, R., Brix, A., 2009. Special Engineering Aspects. Eibl R, Eibl D, Pörtner R, Catapano G, Czermak P (eds). Cell and Tissue Reaction Engineering: Principles and Practice. Springer-Verlag Berlin. Germany; pp 106-120.

De-Leeuw, O.S., Peeters, B., 1999. Complete nucleotide sequence of Newcastle disease virus: evidence for the existence of a new genus within the subfamily Paramyxovirinae. J. Gen. Virol., 80, 131-136.

El-Yuguda, A., Baba, S.S., Geidam, Y.A., 2014. Specific antibody response of village chickens to single or combined Newcastle disease and infectious bursal disease vaccines. J. Adv. Vet. Anim. Res., 1, 16-20.

Gohm, D.S., Thur, B., Hofmann, M.A., 2000. Detection of Newcastle disease virus in organs and feces of experimentally infected chickens using RT-PCR. Avian Pathol., 29, 143-152.

Hassan, W., Khair, S.A.M., Mochotlhoane, B., Abolnik, C., 2010 Newcastle disease outbreaks in the Sudan from 2003 to 2006 were caused by viruses of genotype 5d. Virus Genes, 40, 106-110.

Islam, M.A., Islam, M.A., Hossain, M.T., Sadekuzzaman, M., Hossain, M.G., Hossain, M.M., 2003. Isolation and Identification of Neurtropic Newcastle Disease Virus from Naturally Infected Fayoumi Chickens of Bangladesh. Bangl. Vet. J., 37, 1-9.

Kafi, M.A., Rahman, M.B., Amin, M.M., Islam, M.R., Rahman, M.M., Rahman, M.K., 2003. Comparative Serological Responses and Protection Conferred by Vaccination with V4HR and BCRDV in Chickens. Bangl. J. Vet. Med., 1, 25-27

Kho, C.L., MohdAzmi, M.L., Arshad, S.S., Yusoff, K., 2000. Performance of an RT-nested PCR and ELISA for detection of Newcastle disease virus. J. Virol. Methods, 86, 71-83.

King, D.J., Seal, B.S., 1997. Biological and molecular characterization of Newcastle disease virus isolates from surveillance of live bird markets in the Northeastern United States. Avian Dis., 41, 683-689.

Lamb, R.A., Kolakofsky, D., 2001. Paramyxoviridae: the viruses and their replication. In: Knipe DM and PM Howley (eds), Fields Virology, 4th edn, pp. 1305–1340. Lippincott Williams & Wilkins, Philadelphia.

Liu, H., Wang, Z., Wang, Y., Sun, C., Zheng, D., Wu, Y., 2008. Characterization of Newcastle disease virus isolated from Waterfowl in China. Avian Dis., 52, 150-155.

Mathivanan, K., Kumanan, K., Nainar, A.M., 2004.

Characterization of Newcastle disease virus isolated from apparently normal guinea fowl (Numida meleagris). Vet. Res. Commun., 28, 171-177.

Mayo, M.A., 2002. A summary of the taxonomic changes recently approved by ICTV. Arch. Virol., 147, 1655-1656.

Meulemans, G., van denBerg, T.P., Decaesstecker, M., Boschmans, M., 2002. Evolution of pigeon Newcastle disease virus strains. Avian Pathol., 31, 515-519.

Molia, S., Samaké, K., Diarra, A., Sidibé, M.S., Doumbia, L., Camara, S., Kanté, S., Kamissoko, B., Diakité, A., Gil, P., Hammoumi, S., de Almeida, R.S., Albina, E., Grosboisa, V., 2011. Avian influenza and Newcastle disease in three risk areas for H5N1 highly pathogenic avian influenza in Mali, 2007-2008. Avian Dis., 55, 650-658.

Murmu, R., Islam, M.N., Juli, M.S.B., Khan, M.A.S., Harun-ur-Rashid, S., Hossain, F.M.A., Rahman, M.M., 2014. Pathogenicity and immunosuppresive properties of GM-97 strain of infectious bursal disease virus in commercial broiler chicken. J. Adv. Vet. Anim. Res., 1, 1-7.

Nanthakumar, T., Kataria, R.S., Tiwari, A.K., Butchaiah, G., Kataria, J.M., 2000. Pathotyping of Newcastle disease viruses by RT-PCR and restriction enzyme analysis. Vet. Res. Commun., 24, 275-286.

Nasrin, M., Khan, M.Z.I., Siddiqi, M.N.H., Masum, M.A., 2013. Mobilization of immunoglobulin (Ig)-containing plasma cells in Harderian gland, cecal tonsil and trachea of broilers vaccinated with Newcastle disease vaccine. Tissue Cell, 45, 191-197.

Nooruzzaman, M., Mazumder, A.C., Khatun, S., Chowdhury, E.H., Das, P.M., Islam, M.R., 2013. Pathotypic and genotypic characterization of two Bangladeshi isolates of Newcastle Disease Virus of chicken and pigeon origin. Transbound. Emerg. Dis., doi: 10.1111/tbed.12086.

Pazhanivel, N., Balsubramaniam, G.A., George, V.T., Mohan, B., 2002. Study of natural outbreak of Newcastle disease in and around Namakkal. Indian Vet. J., 79, 293-294.

Peeters, B.P., de Leeuw, O.S., Koch, G., Gielkens, A.L., 1999. Rescue of Newcastle disease virus from cloned cDNA: evidence that cleavability of the fusion protein is a major determinant for virulence. J. Virol., 73, 5001-5009.

Rahman, M., Rahman, M.B., Sarkar, S.K., Khan, M.F.R., Nazir, K.H.M.N.H., 2004. Comparative efficacy of experimentally prepared V4HRND vaccine with commercial V4HR Newcastle disease vaccine and BCRDV vaccine. Prog. Agric., 15, 97-102.

Reddy, G.S., Srinivasan, V.A., 1997. Evaluation of BHK-21 cell-line adapted Newcastle disease vaccine. Indian J. Anim. Sci., 67, 357-358.

Saha, S., Islam, M.A., Rahman, M.M., Alam, K.M.T., 1997. Biological characters of the field isolates of Newcastle disease virus. Bangl. Vet. J., 31, 123-125.

Steneroden, K., 2004. Newcastle Disease. The Center for Food Security and Public Health: Iowa State University.

Stephen, W., Rao, S.B.V., Agarwal, K.K., 1975. Standard methods for the examination of avian pathogens. Fed. Reg., 42, 434.

Talha, A.F.S.M., Hossain, M.M., Chowdhury, E.H., Bari, A.S.M., Islam, M.R., Das, P.M., 2001. Poultry diseases occurring in Mymensingh district of Bangladesh. Bangl. Vet., 18, 20–23.

Published

2014-06-29

How to Cite

Rahman Khan, M. F. ., Ahmed, S. ., Liakot Hossen, M. ., A. Imran, M., Nazmul Hussain Nazir, K. H. M. ., Rahman, M. ., T. Rahman, M., & B. Rahman, M. (2014). Isolation, molecular detection and BHK-21 adaptation of Newcastle disease virus of field cases in layer farms of Bangladesh. Scientific Journal of Microbiology, 3(6), 66-73. Retrieved from https://www.sjournals.com/index.php/sjm/article/view/1367

Issue

Section

Original Article