Open Access
Copyright: The authors. This article is an open access
article licensed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/2.0) which permits unrestricted use,
distribution and reproduction in any medium, provided the work is properly
cited.
R esearch
(Published online:
04-05-2015)
2.
Phylogenetic analysis of Dichelobacter
nodosus serogroup-specific fimA gene from ovine footrot
in Andhra Pradesh -
N. Vinod Kumar, A.
Karthik, S. Vijayalakhsmi and D. Sreenivasulu
Veterinary World, 8(5): 567-571
doi:
10.14202/vetworld.2015.567-571
N.
Vinod Kumar:
Department of Veterinary Microbiology, College of Veterinary
Science, Sri Venkateswara Veterinary University,
Tirupati - 517 502, Andhra Pradesh, India; nagaram_vinod@yahoo.com
A.
Karthik:
Department of Veterinary Microbiology, College of Veterinary
Science, Sri Venkateswara Veterinary University,
Tirupati - 517 502, Andhra Pradesh, India; akmicro7@gmail.com
S.
Vijayalakhsmi:
Department of Veterinary Microbiology, College of Veterinary
Science, Sri Venkateswara Veterinary University,
Tirupati - 517 502, Andhra Pradesh, India; vijaya.sidd@gmail.com
D.
Sreenivasulu: Department of Veterinary Microbiology, College
of Veterinary Science, Sri Venkateswara Veterinary University,
Tirupati - 517 502, Andhra Pradesh, India;
dsreenivasulu10@gmail.com
Received: 27-12-2014, Revised: 24-03-2015, Accepted: 30-03-2015,
Published online: 04-05-2015
Corresponding author:
N. Vinod Kumar, e-mail: nagaram_vinod@yahoo.com
Citation: Vinod Kumar
N, Karthik A, Vijayalakhsmi S, Sreenivasulu D (2015) Phylogenetic
analysis of Dichelobacter nodosus serogroup specific
fimA gene from ovine footrot in Andhra Pradesh, Veterinary
World 8(5):567-571.
Abstract
Aim:
Identification of different serogroups of Dichelobacter
nodosus prevailing in the region and to understand the degree
of genetic heterogeneities among the different isolates of D.
nodosus.
Materials and Methods: A total of 150 exudate samples of
footrot lesions with a lesion score of 2-4 were collected from
naturally infected sheep. The samples were screened by polymerase
chain reaction (PCR) targeting D. nodosus specific 16srRNA.
Of 150 samples screened, 70 samples were found to be positive. The
positive samples were attempted for isolation of D. nodosus,
out of which 16 isolates were recovered. All the isolates were
subjected to serogrouping by multiplex PCR targeting fimA
gene using A-I serogroup specific primers.
Results: Of 16 isolates, 7 (43.75%) isolates were serogroup B,
4 (25.00%) isolates were serogroup A, 3 isolates (18.75%) were
serogroup I and 2 (12.5%) isolates yielded both serogroup A and B.
phylogenetic analysis was performed using neighbor-joining
algorithm of the ClustelX2 software in order to study whether the
serogroups isolated in the present investigation differed
genetically from other published serogroups. The fimA gene
sequence of present isolates of serogroups A, B, and I were
segregated into three distinct groups with high bootstrap values.
The serogroup B clustered with Australian isolate of serotype B1
suggesting high genetic similarity of the present isolate with
serotype B1.
Conclusions: The clinical samples were collected from
suspected outbreaks of footrot and identified the prevalence of
D. nodosus by PCR targeting 16srRNA gene. Identified
serogroups A, B, and I from different districts of Andhra Pradesh.
The phylogenetic analysis will help for the tentative
identification of serotypes present in the serogroup and to
understand the degree of genetic heterogeneities among the
different isolates of D. nodosus.
Keywords: fimA gene, -footrot,-
phylogenetic analysis, polymerase chain reaction.
References
1. Thomos, J.H. (1962) The differential diagnosis of footrot
in sheep. Aust. Vet. J., 38: 159-163.
http://dx.doi.org/10.1111/j.1751-0813.1962.tb16034.x |
|
|
2. Claxton, P.D., Ribeiro, L.A. and Egerton, J.R. (1983)
Classification of Bacteroides nodosus by agglutination tests.
Aust. Vet. J., 60(11): 331-334.
http://dx.doi.org/10.1111/j.1751-0813.1983.tb02834.x |
|
|
3. La Fontaine, S., Egerton, J.R. and Rood, J.I. (1993)
Detection of Dichelobacter nodosus using species-specific
oligonucleotides as PCR primers. Vet. Microbiol., 35(1-2):
101-107.
http://dx.doi.org/10.1016/0378-1135(93)90119-R |
|
|
4. Dhungyel, O.P., Whittington, R.J. and Egerton, J.R. (2002)
Serogroup specific single and multiplex PCR with pre-enriched
culture and immuno-magnetic bead capture for identifying
strains of D.nodosus in sheep with footrot prior to
vaccination. Mol. Cell. Probes., 16(4): 285-296.
http://dx.doi.org/10.1006/mcpr.2002.0427
PMid:12270270 |
|
|
5. Nieuwhof, G.J. and Bishop, S.C. (2005) Costs of the major
endemic diseases of sheep in Great Britain and the potential
benefits of reduction in disease impact. Anim. Sci., 81:
23-29.
http://dx.doi.org/10.1079/ASC41010023 |
|
|
|
6. Stewart, D.J. (1989) Footrot in sheep. In: Egerton, J.R.,
Yong, W.K., Yong, W.K., Riffikin, G.G., editors. Footrot and
Foot Abscesses of Ruminants. C.R.C. Press, Boca Raton, FL.
p5-45. |
|
|
7. Hussain, I., Wani, S.A., Qureshi, S.D. and Farooq, S.
(2009) Serological diversity and virulence determination of
Dichelobacter nodosus from footrot in India. Mol. Cell.
Probes., 23(2): 112-114.
http://dx.doi.org/10.1016/j.mcp.2009.01.003
PMid:19272343 |
|
|
|
8. Farooq, S., Wani, S.A., Hussain, I. and Bhat, M.A. (2010)
Prevalence of ovine footrot in Kashmir, India and molecular
characterization of Dichelobacter nodosus by PCR. Indian J.
Anim. Sci., 80: 826-830. |
|
|
9. Rather, M.A., Wani, S.A., Hussain, I., Bhatt, M.A., Kabli,
Z.A. and MAgray, S.N. (2011) Determination of prevalence and
economic impact of ovine foortot in cental Kashmir India with
isolation and molecular characterization of Dichelobacter
Nodosus. Anaerobe, 17(2): 73-77.
http://dx.doi.org/10.1016/j.anaerobe.2011.02.003
PMid:21397709 |
|
|
|
10. Sreenivasulu, D., Vijayalakshmi, S., Rani Prameela, D.,
Karthik, A., Wani, S.A. and Hussain, I. (2013) Prevalence of
ovine foortot in the tropical climate of southern India and
isolation and characterisation of Dichelobacter nodosus. Sci.
Technol. Rev. OIE., 32(3): 869-877. |
|
|
|
11. Vinod Kumar, N., Karthik, A., Sreenivasulu, D. and Vijaya
Bhaskar Reddy, M. (2013) Incidence of footrot in sheep during
summer at Thottambedu and B.N. Kandriga mandals of Chittoor
District, Andhra Pradesh. Int. J. Adv. Sci. Technol. Res.
3(4): 248-256??? . |
|
|
|
12. Thorley, C.M. (1976) A simplified method for the isolation
of bacteroides nodusus from ovine foot-rot and studies on its
colony morphology and serology. J. Appl. Microbiol., 40(3):
301-309. |
|
|
|
13. Hall, T.A. (1999) BioEdit: A user friendly biological
sequence alignment editor and analysis programme for Windows
95/98/NT. Nucleic Acids Symp Ser., 41: 95-98. |
|
|
14. Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F.
and Higgins, D.G. (1997) The Clustal X windows interface:
Flexible strategies for multiple sequence alignment aided by
quality analysis tools. Nucleic Acids Res., 25(24): 4876-4882.
http://dx.doi.org/10.1093/nar/25.24.4876 |
|
|
15. Felsenstein, J. (1985) Confidence limits on phylogenies:
An approach using boot strap. Evolution, 39(4): 783-791.
http://dx.doi.org/10.2307/2408678 |
|
|
16. Egerton, J.R., Ghimire, S.C., Dhungyel, O.P., Shresta,
H.K., Joshi, H.D., Joshi, B.R., Abboff, K.A. and Kristo, C.
(2002) Eradication of virulent footrot from sheep and goats in
an endemic area of Nepal and an evaluation of specific
vaccination. Vet. Rec., 151(10): 290-295.
http://dx.doi.org/10.1136/vr.151.10.290 |
|
|
17. Gurung, R.B., Dhungyel, O.P., Tshering, P. and Egerton,
J.R. (2006) The use of an autogenous D.nodosus vaccine to
eliminate clinical signs of virulent footrot in sheep flock in
Bhutan. Vet. J., 172(2): 356-363.
http://dx.doi.org/10.1016/j.tvjl.2005.04.032
PMid:16019243 |
|
|
18. Bhatt, M.A., Wani, S.A., Muzafar, M., Rather, M.A., Taku,
A.K. and Khandey, F. (2013) Non - Specificity of primers used
for PCR based serogrouping Dichelobacter nodosus and
identification of novel D. Nodosus strain. Anaerobe., 21:
58-61.
http://dx.doi.org/10.1016/j.anaerobe.2013.03.009
PMid:23542117 |
|
