1994-2019. All Rights Reserved. Online Journal of
Veterinary Research. You may not store these pages in any form except
for your own personal use. All other usage or distribution is illegal under
international copyright treaties. Permission to use any of these
pages in any other way besides the before mentioned must be gained in writing
from the publisher. This article is exclusively copyrighted in its
entirety to OJVR publications. This article may be copied once but may not be,
reproduced or re-transmitted without the express permission of the editors.
OJVRTM
Online Journal of Veterinary Research©
Established 1994
ISSN 1328-925X
Volume 22
(9):832-846, 2018.
Detection methods for Mycobacterium avium paratuberculosis in goats
Forough Zarei Kordshouli1, Azizollah
Khodakaram Tafti1, Bita
Geramizadeh2, Masoud Haghkhah3
1Department(s) of
Pathology, 3Microbiology, School of Veterinary Medicine, Shiraz University, 2Pathology,
Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
ABSTRACT
Kordshouli FZ, Tafti AK, Geramizadeh B, Haghkhah M., Detection methods for Mycobacterium avium paratuberculosis
in goats, Onl J Vet Res., 22 (9):832-846, 2018. We describe molecular, acid
fast staining, histopathology and cultivation methods for diagnosis of Mycobacterium avium
paratuberculosis in goats. Ileum, cecum, colon and mesenteric
lymph nodes were excised from 30 goats suspected of paratuberculosis. Hematoxylin and eosin and acid fast stainings were performed for histopathological and
histochemical examinations. For MAP isolation, Middlebrook
7H9 broth-based associated with OADC (oleic acid, albumin, dextrose, and
catalase) supplement with/without mycobactin J were
used as a medium.
In order to cultivation, the double incubation method was used for tissue
decontamination. IS900 specific DNA probe
detection by the polymerase chain reaction
was performed from broth based media and DNA extractions of tissue speciemens. MAP was cultured after 3-10 days in Middlebrooke 7H9 medium with and without mycobactin J from tissue samples of 15 suspected goats.
Bacterial culture revealed all of 15 goats were infected with MAP. Acid-fast
staining of broth based cultures
were positive in 47% of ileums, 60% of cecums, 33% of
colons and 40% of mesenteric lymph nodes with mycobactin
J and in 60% of ileums, 53% of cecums, 40% of colons
and 33% of mesenteric lymph nodes without mycobactin
J. MAP specific IS900 gene with 413bp was demonstrated in 70% of intestinal
tissue specimens and in all of the
samples in both of with and without mycobactin J
cultures. Histopathological examination revealed diffuse granulomatous enteritis
and lymphadenitis. Acid fast staining of these lesions showed two different paucibacillary or multibacillary
lesions. Positive acid fast staining was observed in 47%
and 43% of intestinal tissue specimens as paucibacillary and multibacillary, respectively. Multifocal granulomatous lymphadenitis associated with caseous necrosis and calcification were
observed in the mesenteric lymph nodes. The results of this study showed
culture of tissue samples with or without mycobactin
J associated with IS 900 PCR can detect MAP in almost all of affected goats to paratuberculosis.
Keywords: IS900 PCR, Culture, Histopathology, Acid fast, Johne’s
disease, Goat.
1994-2019. All Rights Reserved. Online Journal of
Veterinary Research. You may not store these pages in any form except
for your own personal use. All other usage or distribution is illegal under
international copyright treaties. Permission to use any of these
pages in any other way besides the before mentioned must be gained in writing
from the publisher. This article is exclusively copyrighted in its
entirety to OJVR publications. This article may be copied once but may not be,
reproduced or re-transmitted without the express permission of the editors.
OJVRTM
Online Journal of Veterinary Research©
Established 1994
ISSN 1328-925X
Volume 22 (9):832-846, 2018.
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