Isolation of Mycobacterium avium subsp. paratuberculosis from mouflon in Bulgaria

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Abstract

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of paratuberculosis (John’s disease) mainly in large and small domestic and wild ruminants, and suspected causative agent in human Crohn’s disease. In Bulgaria, paratuberculosis is still poorly researched in both groups of ruminants. We present results of the first in-depth study of mouflon, grown free in one hunting reserve in the Western region of the country. The aim was to prove the presence of MAP in diagnostic materials from regularly hunted or dead mouflon suspected for paratuberculosis. Small intestine and mesenteric lymph nodes (MLN) from 12 hunted and 4 dead mouflon and 10 faecal samples (Fc) were studied in the period of 2009–2013. Typical for paratuberculosis pathomorphological lesions were observed in four mouflon (of 16 examined). The intestinal wall was thickened, strongly folded and soft, with severe hyperemia. The MLN were enlarged, soft, with marbled appearance. The affected section of the ileum showed hyperplasia of the mucous corion and submucosa with diffuse infiltration of epithelioid cells. Lymphadenopathy with atrophy of T and B lymphocytes areas was observed in the mesenteric lymph nodes. For bacteriological isolation of MAP, the tissue and faecal samples were decontaminated with NALC-NaOH, cultured in Middlebrook 7H9 Broth and on Herrold’s medium. The Ziehl–Neelsen stained smears and isolates were examined microscopically for acid-fast bacteria. Presence of MAP was observed in tissue samples of 4 (25%) mouflon and in 2 (20%) faecal samples. The same samples were confirmed by the IS900 PCR for the presence of specific for MAP fragments with a commercial amplification kit. The cases of paratuberculosis found at different times in the free-living mouflon in our study prove that the disease exists in Bulgaria and highlight the need for more serious control of the disease among wild and domestic ruminants.

About the authors

T. Savova

National Diagnostic and Research Veterinary Medical Institute

Author for correspondence.
Email: fake@neicon.ru

Associate Professor, PhD (Veterinary Medicine), Head of National Reference Laboratory for Animal Tuberculosis

Sofia

Bulgaria

R. Petrova

National Diagnostic and Research Veterinary Medical Institute

Email: fake@neicon.ru

Assistant Professor, PhD (Veterinary Medicine), Head of the National Reference Laboratory for Anthrax and Rabies in Animals, Department of Pathomorphology

Sofia

Bulgaria

V. Valcheva

The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences

Email: violeta_valcheva@mail.bg

Violeta Valcheva, Assistant Professor, PhD (Biology), Department of Infectious Microbiology, Laboratory of Zoonosis and Bacterial Virulence

26, Acad. Georgi Bonchev str., Sofia 1113

Phone:+3592 979-31-57

Bulgaria

M. Bonovska

The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences

Email: m_bonovska@abv.bg

Magdalena Bonovska, Professor, PhD (Veterinary Medicine), Department of Infectious Microbiology, Laboratory of Zoonosis and Bacterial Virulence

26, Acad. Georgi Bonchev str., Sofia 1113

Phone:+3592 979-31-57

Bulgaria

H. Najdenski

The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences

Email: fake@neicon.ru

Corresponding Member, Professor, DS (Veterinary Medicine), Director, Head of Department of Infectious Microbiology and Laboratory of Zoonosis and Bacterial Virulence

Sofia

Bulgaria

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Copyright (c) 2020 Savova T., Petrova R., Valcheva V., Bonovska M., Najdenski H.

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