Determination of prevalence, serological diversity, and virulence of Dichelobacter nodosus in ovine footrot with identification of its predominant serotype as a potential vaccine candidate in J&K, India

  • Shakil A. Wani
  • S. FarooqEmail author
  • Z. A. Kashoo
  • I. Hussain
  • M. A. Bhat
  • M. A. Rather
  • S. Aalamgeer
Regular Articles


The aim of this study was to determine the prevalence, serological diversity, and virulence of Dichelobacter nodosus in footrot lesions of sheep and identification of its predominant serotype as a potential vaccine candidate. The overall prevalence of footrot in sheep was 16.19%, and ranged from 13.69 to 19.71%, respectively. A total of 759 flocks with 22,698 sheep were investigated for footrot and 2374 clinical samples were collected from naturally infected sheep exhibiting footrot lesions. Of the 2374 samples collected, 1446 (60.90%) were positive for D. nodosus by polymerase chain reaction (PCR). These positive samples when subjected to serogroup-specific multiplex PCR, 1337 (92.46%) samples carried serogroup B, 247 (17.08%) possessed serogroup E, 86 (5.94%) serogroup I, and one (0.069%) serogroup G of D. nodosus. While mixed infection of serogroups B and E was detected in 127 (8.78%), B and I in 46 (3.18%) and B, E, and I in 26 (1.79%) samples, respectively. The serogroup B of D. nodosus was the predominant (92.47%) serogroup affecting sheep population with footrot followed by serogroup E (19.91%) and serogroup I (4.57%), respectively. Virulent status of D. nodosus strains were confirmed by presence of virulence-specific integrase A (intA) gene and the production of thermostable proteases. The intA gene was detected in 709 (72.79%) samples while gelatin gel test carried out on 246 representative isolates all positive for intA gene produced thermostable proteases, confirming their virulence nature. The PCR-restriction fragment length polymorphism (PCR-RFLP) of whole fimA gene of serogroup B revealed the predominance of serotype B5 (82.97%) of serogroup B. This information suggests that serotype B5 is the predominant serotype of D. nodosus associated with severe footrot lesions in sheep in Jammu & Kashmir (J&K), India. Hence, this serotype can be a potential vaccine candidate for the effective control and treatment of ovine footrot.


Ovine footrot Prevalence intA gene fimA gene Gelatin gel test 



The authors acknowledge Dr. O.P. Dhungyel, University of Sydney, Camden, NSW 2570, Australia, for the supply of serogroup-specific positive control DNA for the study.

Funding information

The Niche Area of Excellence (NAE)- ICAR, New Delhi.) financially assisted the study.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest.

Ethic statement

This study was performed with the permission of the Institutional Animal Ethics Committees (Number AU/FVS/IAEC-12/83-85) of Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, J&K, India.


  1. Bhat, M.A., Wani, S.A., Hussain, I., Magray, S.N., Muzafar, M. (2012). Identification of two new serotypes within serogroup B of Dichelobacter nodosus. Anaerobe 18, 91–95.CrossRefGoogle Scholar
  2. Cheetham, B.F., Tanjung, L.R., Sutherland, M., Druitt, J., Green, G., McFarlane, J., Bailey, G.D., Seaman J.T. and Katz, M.E. (2006). Improved diagnosis of virulent ovine footrot using the intA gene. Vet. Microbiol. 116 (1–3), 166–174.CrossRefGoogle Scholar
  3. Chetwin, D.H., Whitehead, L.C. and Thorley, S.E. (1991). The recognition and prevalence of Bacteriodes nodosus serotype M in Australia and New Zealand. Australian Veterinary Journal 68, 154–155.CrossRefGoogle Scholar
  4. Claxton, P.D. (1986). Serogrouping of Bacteroides nodosus isolates. In: Proceedings of a Workshop, Melbourne, 1985, CSIRO, Division of Animal Health and Australian Wool Corporation, Glebe, Sydney, N.S.W, Australia. 131–134.Google Scholar
  5. Claxton, P.D., Ribeiro, L.A and Egerton, J.R. (1983a).Classification of Bacteroides nodosus by agglutination test. Aust. Vet. J. 60 (11), 331–334. CrossRefGoogle Scholar
  6. Claxton, P.D., Ribeiro, L.A. and J.R. Egerton. (1983b). Classification of Bacteroides nodosus by agglutination test. Aust. Vet. J. 70, 123–126.Google Scholar
  7. Dhungyel, O.P., Whittington, R.J. & Egerton, J.R. (2002). Serogroup specific single and multiplex PCR with pre-enrichment 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.CrossRefGoogle Scholar
  8. Every, D., and Skerman, T.M. (1982). Protection of sheep against experimental footrot by vaccination with pili purified from Bacteroides nodosus. New Zealand Veterinary Journal 30, 156–158.CrossRefGoogle Scholar
  9. Farooq, S., Wani, S.A., Hussain, I. & Bhat, M.A. (2010). Prevalence of ovine footrot in Kashmir, India and molecular characterization of Dichelobacter nodosus by PCR. Indian J. Anim. Sci. 80 (9), 826–830.Google Scholar
  10. Fontaine, LA., Egerton, J.R. & Rood, J.I. (1993). Detection of Dichelobacter nodosus using species-specific oligonucleotides as PCR primers. Vet. Microbiol. 35 (1–2), 101–117.CrossRefGoogle Scholar
  11. Ghimire, S.C., Egerton, J.R., Dhungyel, O.P., & Joshi, H.D. (1998). Identification and characterization of serogroup M among Nepalese isolates of Dichelobacter nodosus, the transmitting agent of footrot in small ruminants. Vet. Microbiol. 62 (3) 217–233.CrossRefGoogle Scholar
  12. Glynn, T. (1993). Benign footrot-An epidemiological investigation into the occurrence of effects on production, response to treatment and influence of environment factors. Aust. Vet. J. 70, 7–12.CrossRefGoogle Scholar
  13. Gradin, J.L., Sonn, A.E., Petrovska, L. (1993). Serogrouping of Bacteroides nodosus isolates from 62 sources in the United States. Am. J Vet. Res. 54, 1069–1073.Google Scholar
  14. Grogono, T.R. & Johnston, A.M. (1997). A study of ovine lameness, MAFF Open Contract OC 5945 K, DEFRA Publications, London.Google Scholar
  15. Gurung, R.B., Tshering, P., Dhungyel, O.P. & Egerton, J.R. (2006). Distribution and prevalence of footrot in Bhutan. Vet. J. 171(2), 346–51.CrossRefGoogle Scholar
  16. Hindmarsh, F.& Fraser, J. (1985). Serogrouping of Bacteroides nodosus isolated from ovine footrot in Britain. Vet. Rec. 116 (7), 187–88.Google Scholar
  17. Kingsley, D.F, Hindmarsh, F.H, Liardet, D.M., & Chetwin, D.H. (1986). Distribution of serogroups of Bacteroides nodosus with particular reference to New Zealand and the United Kingdom. Footrot in Ruminants. (Eds) Stewart D J, Peterson J E, Mckern N M, Emery and D L. Published by CSIRO/Australian wool corporation, Sydney, N.S.W. Australia. 143–46.Google Scholar
  18. Marshall, D.J., Walker, R.I., Cullis, B.R., Luff, M.F. (1991). The effect of footrot on body weight and wool growth of sheep. Aust. Vet. J. 68, 45–49.CrossRefGoogle Scholar
  19. Mc Kern, N.M., Elleman, T.C. and Hoyne, P.A. (1989). Molecular basis of antigenic variation in pili (fimbriae) of Bacteroides nodosus - application to footrot vaccines. In: Footrot and Foot Abscess of Ruminants (Eds. J.R. Egerton, W.K. Yong and G.G Riffkin). CRC Press, Boca Raton, FL.Google Scholar
  20. Palmer, M.A. (1993). A gelatin test to detect activity and stability of proteases produced by Dichelobacter (Bacteroides) nodosus. Vet. Microbiol. 36 (1–2),113–122.CrossRefGoogle Scholar
  21. Rather, M.A., Wani, S.A, Hussain, I., Bhat, M. A., Kabli, Z. A. and Magray, S.N. (2011). Determination of prevalence and economic impact of ovine footrot in Central Kashmir India with isolation and molecular characterization of Dichelobacter nodosus. Anaerobe 17, 73–77.CrossRefGoogle Scholar
  22. Sreenivasulu, D., Vijayalakshmi, S., Raniprameela, D., Karthik, A., Wani, S.A & Hussain, I.(2013). Prevalence of ovine footrot in the tropical climate of southern India and isolation and characterisation of Dichelobacter nodosus. Rev. sci. tech. Off. int. Epiz., 32 (3).Google Scholar
  23. Stewart, D. (1989). Footrot in sheep. In: Egerton JR, Yong WK, Riffkin GG, editors. Footrot and Foot Abscesses of Ruminants, CRC press, Boca Raton, Florida. 5–45.Google Scholar
  24. Stewart, D.J. (1978).The role of various antigenic factors of Bacteroides nodosus in eliciting protection against foot rot in vaccinated sheep. Research in Veterinary Science 24, 14–19.CrossRefGoogle Scholar
  25. Stewart, D.J., Claxton, P.D., 1993. Ovine foot rot: clinical diagnosis and bacteriology. In: Corner, L.A., Bagust, T.J. (Eds.), Australian Standard Diagnostic Techniques for Animal Diseases. CSIRO Publications, Victoria, Australia, pp. 1–27.Google Scholar
  26. Wani, S.A., Samanta, I., Buchh, A.S. & Bhat, M.A. (2004). Molecular detection and characterization of Dichelobacter nodosus in ovine footrot in India. Mol. Cell. Probes 18 (5), 289–291.CrossRefGoogle Scholar
  27. Wani, S.A., Samanta, I., Shah, S.S. (2007). Isolation and characterization of Dichelobacter nodosus from ovine and caprine footrot in Kashmir, India. Res Vet Sci 83, 141–4.CrossRefGoogle Scholar
  28. Wassink, G.J., Grogono, R. Thomas, Moore, L.J. & Green, L.E. (2003). Risk factors associated with the prevalence of footrot in sheep from 1999 to 2000. Vet. Rec. 152 (12), 351–58.CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Bacteriology Laboratory, Division of Veterinary Microbiology and ImmunologySKUAST-K, Shuhama (Alusteng)SrinagarIndia
  2. 2.Division of Veterinary Microbiology and ImmunologySher-e-Kashmir University of Agricultural Sciences and Technology of Jammu (SKUAST-J)JammuIndia
  3. 3.Department of Sheep HusbandryGovernment of Jammu and KashmirJammuIndia

Personalised recommendations