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Open Access
Copyright: The authors. This article is an open access
article licensed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/2.0) which permits unrestricted use,
distribution and reproduction in any medium, provided the work is properly
cited.
Research (Published
online : 26-08-2013)
17. Investigation on the status of Johne's disease based on agar gel
immunodiffusion, ziehl-neelsen staining and nested PCR approach in two cattle
farm - Anand Mohan, Pranabananda Das, Neelam Kushwaha, Kaliaperumal
Karthik and Ankush Kiran Niranjan
Veterinary World, 6(10): 778-784
doi:
10.14202/vetworld.2013.778-784
Abstract
Background and Methods:
Paratuberculosis is a chronic disease of ruminant, caused by
Mycobacterium avium subsp. paratuberculosis (Map), clinically
infected animals produce high level of antibodies in blood and
shed detectable amount of Map organisms in feces. Several
serological and molecular tests are utilized for detection of
antibodies or DNA of the organism in clinical samples. Present
study indicates the status of paratuberculosis in two distinct
cattle farms with different organizational set-ups viz. organized
and unorganized. We used agar gel immunodiffusion (AGID) assay for
the detection of antibodies in blood. Ziehl-Neelsen (ZN) staining
of fecal smears was done to observe acid-fast bacilli and Nested
PCR targeted to IS900 and f57 sequences, was performed to confirm
the pathogen.
Results: Sera samples of cattle, from organized farm, did
not show any visible precipitating band with AGID assay. However,
fecal smears of few cattle (3.57%) were positive for acid-fast
bacilli. When confirmed with nested PCR, only one fecal sample
(0.71%) was found positive for Map. In case of unorganized farm, a
large number of cattle (38.75%) showed precipitating antibodies
with AGID assay and the percentage of fecal smears that showed
acid-fast bacilli was 26.62%. Nevertheless, fecal samples
containing Map DNA was confirmed in 14.37% of fecal sample by
nested PCR.
Conclusions: An organized farm, with better hygiene and
management practices, showed lesser occurrence of paratuberculosis
in cattle in comparison to unorganized farm. Not all AGID assays
positive cattle might be an efficient shedder of Map and mare
detection of acid-fast bacilli in fecal smears did not always
indicate the presence of Map organism. Cattle infected with JD
were mostly in the age group of six years and above.
Keywords: AGID, f57, Johne's disease, IS900, Nested PCR,
Ziehl-Neelsen staining, Mycobacterium avium subspecies
paratuberculosis.
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