Documenti di Didattica
Documenti di Professioni
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50 (1): 145-149
ISSN 1110-2047, www.alexjvs.com
DOI: 10.5455/ajvs.230284
ABSTRACT
Key words: This study reports successful isolation and characterization of velogenic genotype VI Newcastle disease
Newcastle
disease virus;
Velogenic;
pigeon;
Genotype IV;
Egypt
virus (NDV) from pigeons in Egypt. A pool of brain sample was collected from three pigeons showing
torticollis and whitish green diarrhea in Desouk, Kafr-elshiekh Province, Egypt, during January, 2014.
Virus Isolation has been pursued by inoculation of brain homogenate into the allantoic sac of 9 day old
specific pathogen free (SPF) embryonated chicken eggs. Identification of the isolate was carried out by
haemagglutination inhibition test using specific polyclonal sera against NDV, RT-PCR and confirmed by
sequencing. Partial deduced amino acid sequence of the fusion protein at the cleavage site revealed that
the isolate possessed the motif 112 K-R-Q-K-R-F117 indicating velogenic entity and phylogenetic analysis
showed clustering of the study sequence with other genotype IV F gene sequences available in GenBank.
The study indicated that pigeons may play epidemiological role in the introduction of a new NDV
genotype VI to commercials chicken, which will increase the epidemiological burden of Newcastle
disease (ND) in Egypt
1.
INTRODUCTION:
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Table (1): Thermo cycler protocol for amplification of partial F gene of NDV
Step
Temperature
Time
Number of cycles
Initial denaturation
RT
50 C
94 C
15 minutes
2 minutes
1cycle
1cycle
Denaturation
94 C
30 seconds
40 cycles
Primer annealing
50 C
30 seconds
Extension
72 C
45 seconds
Final extension
72 C
10 minutes
1cycle
Cooling
4 C
---------
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El-Shazly et al. /Alexandria Journal of Veterinary Sciences 2016, Apr. 49 (1): 145-149
4. CONCLUSION
Isolation of velogenic NDV genotype VI from
pigeons will increase the epidemiological burden of
NDV in Egypt and asses the role of semidomesticated or feral wild birds in prevalence of
new genotypes of NDV however, no recent study
reported the detection of the same genotype from
commercial chicken in Egypt till now therefore,
extensive surveillance of wild birds in addition to
routine surveillance of domestic poultry is a
fundamental requirement.
Acknowledgments
The work team is grateful to all members of
Virology Department, Faculty of Veterinary
Medicine, Cairo University for their excellent
technical assistance.
Fig. 1: Ethidium bromide stained agarose gel electrophoresis containing the RT-PCR products of internal part
of F gene of NDV (320bp), L= DNA ladder (100-3000 bp), CP= Positive control, CN = Negative control,
Lane 2 and 3 = Positive NDV, Lane 1,4 and 5 negative NDV.
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El-Shazly et al. /Alexandria Journal of Veterinary Sciences 2016, Apr. 49 (1): 145-149
Fig. 2: Neighbor-Joining phylogenetic tree for selected NDV sequences from Genotypes (I, II, VI, VII),
rooted to Genotype I and showing the clustering of Study Isolate (black square) with genotype VI NDV
sequences. Tree was generated using MEGA version (7.0.18) with 500 bootstrap replicates.
Submission to GenBank:
Sequence of the isolate (NDV
pigeon/Desouk/Egypt/MS3I/2014) has been
submitted to the GenBank with Accession No.
KR082484
URL:http://www.ncbi.nlm.nih.gov/gquery/?term=
KR082484.
5. REFERENCES
Abdel-Glil, M.Y., Mor, S.K., Sharafeldin, T.A.,
Porter, R.E., Goyal, S.M. 2014. Detection and
characterization of Newcastle disease virus in
formalin-fixed, paraffin-embedded tissues from
commercial broilers in Egypt. Avian Dis. 58:11823.
Alexander, D.J., Manvell, R.J., Kemp, P.A.,
Parsons, G., Collins, M.S., Brockman, S.,
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Abdel-Hamid and Abdel Fattah /Alexandria Journal of Veterinary Sciences 2016, 50 (1): 145-149