Authors:
Gangadhararao Appana Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India
Department of Biotechnology, Acharya Nagarjuna University, Nagarjunanagar, Guntur, Andhra Pradesh, 522510, India

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Dipankar Das Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India

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Maroudam Veerasami Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India

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Ramachandran Lakshmikanthan Senthilkumar Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India

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Munishkumar Durishetty Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India

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B. Ramalakshmi Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India

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Vijay Bahekar Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India

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Falguni Mukherjee Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India

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Dev Chandran Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, 500 032, India

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P. Uday Kumar National Institute of Nutrition, Jamai-Osmania P.O., Hyderabad, India

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B. Sesikeran National Institute of Nutrition, Jamai-Osmania P.O., Hyderabad, India

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Dr. Villuppanoor Alwar Srinivasan Ph.D. National Dairy Development Board, 33, Telecom Nagar, Gachibowli, Hyderabad, 500032, India

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Abstract

A male cattle calf was detected as subclinically and naturally infected with Mycobacterium avium subspecies paratuberculosis (MAP) by a series of antemortem and postmortem tests. The MAP infection was identified by strong antibody and cell-mediated immune (CMI) response by a commercial ELISA kit and an intradermal Johnin test, respectively, in the initial antemortem examination. The antemortem status of the calf was further confirmed by MAP-specific interferon gamma (IFN-γ) response. For detection of IFN-γ response, MAP-specific IFN-γ release assays (IGRAs): (a) immuno capture ELISA (IC-ELISA) and (b) ELISPOT was employed. In addition, the presence of intracellular cytokine IFN-γ was detected by flow cytometry. For all cytokine assays, MAP-specific recombinant antigens HSP65 and 35 kDa were employed to overcome the poor sensitivity and specificity resulting from the use of Johnin, the crude protein purified derivative of MAP. Postmortem examination of the MAP-infected/suspected cattle calf did not reveal any pathognomonic gross lesions in the gastro-intestinal tract. Histopathological examination of multiple organs showed the presence of epithelioid cells/macrophages and edematous lesions in the mesenteric lymph nodes suggestive of MAP; however, no granulomas were observed in the intestinal tract. The necropsy samples of rectum and mesenteric lymph nodes were positive for isolation of MAP by culture in the BACTEC™ MGIT™ 960 system, and acid fast bacilli were demonstrated by fluorescence microscopy confirming the infection. Due to differential and complex expression patterns of MAP antigens reported in literature, a combination of assays such as those based on IGRAs and antibody detection is essential. Therefore, the current experimental evidence confirms the efficacy of the approach adopted. However, further studies will be needed to understand the optimal combination MAP-specific antigens for use in IGRAs or antibody assays that can be used for detecting MAP infection in every stage of the disease.

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Senior editors

Editor(s)-in-Chief: Dunay, Ildiko Rita, Prof. Dr. Pharm, Dr. rer. nat., University of Magdeburg, Germany

Editor(s)-in-Chief: Heimesaat, Markus M., Prof. Dr. med., Charité - University Medicine Berlin, Germany

Editorial Board

  • Berit Bangoura, Dr. DVM. PhD,  University of Wyoming, USA
  • Stefan Bereswill, Prof. Dr. rer. nat., Charité - University Medicine Berlin, Germany
  • Dunja Bruder, Prof. Dr. rer. nat., University of Magdeburg, Germany
  • Jan Buer, Prof. Dr. med., University of Duisburg, Germany
  • Edit Buzas, Prof. Dr. med., Semmelweis University, Hungary
  • Renato Damatta, Prof. PhD, UENF, Brazil
  • Maria Deli, MD, PhD, DSc, Biological Research Center, HAS, Hungary
  • Olgica Djurković-Djaković, Prof. Phd, University of Belgrade, Serbia
  • Jean-Dennis Docquier, Prof. Dr. med., University of Siena, Italy
  • Zsuzsanna Fabry, Prof. Phd, University of Washington, USA
  • Ralf Ignatius, Prof. Dr. med., Charité - University Medicine Berlin, Germany
  • Achim Kaasch, Prof. Dr. med., Otto von Guericke University Magdeburg, Germany
  • Oliver Liesenfeld, Prof. Dr. med., Inflammatix, USA
  • Matyas Sandor, Prof. PhD, University of Wisconsin, USA
  • Ulrich Steinhoff, Prof. PhD, University of Marburg, Germany
  • Michal Toborek, Prof. PhD, University of Miami, USA
  • Susanne A. Wolf, PhD, MDC-Berlin, Germany

 

Dr. Dunay, Ildiko Rita
Magdeburg, Germany
E-mail: ildiko.dunay@med.ovgu.de

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European Journal of Microbiology and Immunology
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European Journal of Microbiology and Immunology
Language English
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