Histopathology: An old yet important technique to diagnose paratuberculosis in non-descript water buffaloes


  • Arbab Sikandar University of Veterinary and Animal Sciences, Lahore, Pakistan
  • Amar Nasir University of Veterinary and Animal Sciences, Lahore, Pakistan
  • Aziz -ur- Rehman University of Veterinary and Animal Sciences, Lahore, Pakistan
  • Hafiz Muhammad Ali University College of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Pakistan
  • Muqadar Shah University of Veterinary and Animal Sciences, Lahore, Pakistan
  • Muhammad Adil University of Veterinary and Animal Sciences, Lahore, Pakistan
  • Imad Khan University of Veterinary and Animal Sciences, Lahore, Pakistan
  • Noor Muhammad Khan University of Veterinary and Animal Sciences, Lahore, Pakistan


Bubalus bubalis, buffaloes, animals, Johne’s disease, histology, gut, serology


Paratuberculosis (PTB) in buffaloes is a chronic enteric disease triggering health implications and huge economic losses in livestock. This study was designed to explore a simple, cost-effective diagnostic approach for PTB in water buffaloes. Blood (5 ml/animal) and intestinal tissue samples accompanied by lymph nodes associated with mesentery were collected from weak and diarrhoeac animals slaughtered at local abattoirs. Out of total n=771 clinically suspected animals, only n=53 carcasses were sampled based on gross observation. Tissue smears of the gut mucosae were obtained and were made adopting special staining protocol. Tissue samples were processed by paraffin sectioning and stained with Ziehl-Neelsen and Hematoxylin-Eosin staining methods. Acid-fast bacilli were observed only in 11/53 cases on mucosal tissue smears. Pressure atrophy of small intestine villi were evident, and the mucosae were found sloughed off. The submucosae were heavily infiltrated with mononuclear cells and multifocal cellular nodules dominated by epithelioid macrophages. The foamy cytoplasm of the macrophages appeared to be engorged with acid fast bacilli and depicted the positive cases. All tissue sections of the suspected samples showed 100% +ve results while only 20.8% samples were found +ve with smear method. All histo-pathologically positive cases were further confirmed by ELISA based serological analysis. Therefore, it was concluded that histopathology is an economical and yet the most trusted tool for diagnosing bubalian PTB in countries like Pakistan.


Download data is not yet available.


Metrics Loading ...


Ali, H.M., A.S. Qureshi, R. Hussain, G. Urbinati, M.Z. Mustafa, F. Ali, A. Manan and L. Massaad-Massade. 2017. Effects of natural environment on reproductive histo-morphometric dynamics of female dromedary camel. Anim. Reprod. Sci., 181: 30-40. DOI: 10.1016/j.anireprosci.2017.03.012

Amemori, T.l., O. Matlova, A. Fischer, W.Y. Ayele, M. Machackova, E. Gopfert and I. Pavlik. 2004. Distribution of Mycobacterium avium subsp. paratuberculosis in the gastrointestinal tract of shedding cows and its application to laproscopic biopsy. Vet. Med. Czech., 49(7): 225-236. DOI: 10.17221/5699-VETMED

Antognoli, M.C., F.B. Garry, H.L. Hirst, J.E. Lombard, M.M. Dennis, D.H. Gould and M.D. Salman. 2007. Characterization of Mycobacterium avium subspecies paratuberculosis disseminated infection in dairy cattle and its association with antemortem test results. J. Vet. Microbiol., 10(3-4): 1016-1025. DOI: 10.1016/j.vetmic.2007.08.017

Bach, E., E.A. Raizman, R. Vanderwal, P. Soto, M. Chaffer, G. Keefe and H. Bach. 2018. Immunogenicity of PtpA secreted during Mycobacterium avium ssp. paratuberculosis infection in cattle. Vet. Immunol. Immunopathol., 198: 1-5. DOI: 10.1016/j.vetimm.2018.02.006

Buergelt, C.D., P.E. Ginn. 2000. The histopathologic diagnosis of subclinical Johne’s disease in North American Bison (Bison bison). Vet. Microbiol., 77(3-4): 325-331. DOI: 10.1016/s0378-1135(00)00317-5

Catton, B. Amber. 2002. Paucibacillary paratuberculosis in a goat. Canadian Vet. J., 43(10): 787-788.
Correa-Valencia, N., Y.M. García-Tamayo and J.A. Fernández-Silva. 2018. Mycobacterium avium subsp. paratuberculosis in Colombia (1924-2016): A review. Rev. Colomb. Cienc. Pec., 31(3): 165-179. DOI: 10.17533/udea.rccp.v31n3a01

Gautam, M., A. Ridler, P.R. Wilson and C. Heuer. 2018. Control of clinical paratuberculosis in New Zealand pastoral livestock. New Zeal. Vet. J., 66(1): 1-8. DOI: 10.1080/00480169.2017.1379914

Gupta, S.K., P.H. Maclean, S. Ganesh, D. Shu, B.M. Buddle, D.N. Wedlock and A. Heiser. 2018. Detection of microRNA in cattle serum and their potential use to diagnose severity of Johne’s disease. J. Dairy Sci., 101(11): 10259-10270. DOI: 10.3168/jds.2018-14785

Jenvey, C.J., J.M. Hostetter, A.L. Shircliff and J.R. Stabel. 2018. Relationship between the pathology of bovine intestinal tissue and current diagnostic tests for Johne’s disease. Vet. Immunol. Immunop., 202: 93-101. DOI: 10.1016/j.vetimm.2018.06.012

Jones, T. Carlyle, R.D. Hunt and N.W. King. 2006. Veterinary Pathology, 6th ed. Blackwell Publishing, USA. p. 498-501.

Maxie, M.G., K.V.F. Jubb, P.C. Kennedy and N.C. Palmer. 2007. Pathology of Domestic Animals, 5th ed. Saunders Elsevier, London, UK. p. 222-225.

Muhammed, I.S., R.A. Tadayon and A.H. Cheema. 1978. Detection of antibodies to Mycobecterium Johnei by Counterimmunoelectrophoresis. Vet. Rec., 102(18): 401-403. DOI: 10.1136/vr.102.18.401

Pavlik, I., Z. Rozsypalova, T. Vesely, J. Bartl, L. Matlova, V. Vrbas, L. Valent, D. Rajsky, I. Mracko, M. Hirko and P. Miskovic. 2000. Control of paratuberculosis in five cattle farms by serological tests and fecal culture during the period 1990-1999. Vet. Med.-Czech, 45(3): 61-70.

Schaik, G.V., M. Pradenas, F.A. Mella, N. Juan and V. Kruze. 2007. Diagnostic validity and costs of pooled fecal samples and individual blood or fecal samples to determine the cow-and herd-status for Mycobacterium avium subsp. paratuberculosis. Prev. Vet. Med., 82(1-2): 159-165. DOI: 10.1016/j.prevetmed.2007.05.018

Sikandar, A. 2018. Histopathology an old yet important technique in modern science, p. 1. Histopathology - An Update, IntechOpen Limited, London, UK.

Sikandar, A., A.H. Cheema, M. Adil, M. Younus, H. Zaneb, M.A. Zaman, M.Y. Tipu and S. Masood. 2013. Ovine paratuberculosis-A histopathological study from Pakistan. J. Anim. Plant Sci., 23(3): 749-753. Available on: http://thejaps.org.pk/docs/v-23-3/12.pdf

Sikandar, A., A.H. Cheema, M. Younus, A. Aslam, M.A. Zaman and T. Rehman. 2012. Histopathological and serological studies on paratuberculosis in cattle and buffaloes. Pak. Vet. J., 32(4): 547-551.

Singh, S.V., A.V. Singh, R. Singh, K.S. Sandhu, P.K. Singh, J.S. Sohal, V.K. Gupta and V.S. Vihan. 2007. Evaluation of highly sensitive indigenous milk ELISA kit with fecal culture, milk culture and fecal-PCR for the diagnosis of bovine Johne’s disease (BJD) in India. Comp. Immunol. Microb., 30(3): 175-186. DOI: 10.1016/j.cimid.2006.12.002

Sivakumar, P., B.N. Tripathi, N. Singh and A.K. Sharma. 2006. Pathology of naturally occurring paratuberculosis in water buffaloes (Bubalus bubalis). Vet. Pathol., 43(4): 455-462. DOI: 10.1354/vp.43-4-455

Uy, M.R.D., J.L. Cruz, M.A. Miguel, M.B.S. Salinas, J.V. Lazaro and C.N. Mingala. 2018. Serological and molecular evaluation of Mycobacterium avium subspecies paratuberculosis (Johne’s disease) infecting riverine-type water buffaloes (Bubalus bubalis) in the Philippines. Comp. Immunol. Microb., 61: 24-29. DOI: 10.1016/j.cimid.2018.11.004

Yoshida, S., T. Araki, T. Asai, K. Tsuyuguchi, K. Arikawa, T. Iwamoto and T. Wada. 2018. Phylogenetic uniqueness of Mycobacterium avium subspecies hominissuis isolated from an abnormal pulmonary bovine case. Infect. Genet. Evol., 62: 122-129. DOI: 10.1016/j.meegid.2018.04.013




How to Cite

Sikandar, A., Nasir, A., Rehman, A. .- ur-., Ali, H. M., Shah, M., Adil, M., … Khan, N. M. (2021). Histopathology: An old yet important technique to diagnose paratuberculosis in non-descript water buffaloes. Buffalo Bulletin, 40(4), 557–564. Retrieved from https://kuojs.lib.ku.ac.th/index.php/BufBu/article/view/3134



Original Article

Most read articles by the same author(s)