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Research
(Published
online: 11-08-2014)
7. Detection of Mycobacterium avium subsp.
paratuberculosis from cattle and buffaloes in Egypt using
traditional culture, serological and molecular based methods
- G. S. Abdellrazeq, M. M. Elnaggar, S. A. Khaliel and A. E.
Gamal-Eldin
Veterinary World, 7(8): 586-593
doi:
10.14202/vetworld.2014.586-593
G. S. Abdellrazeq: Department of Microbiology, Faculty of
Veterinary Medicine, Alexandria University, Edfina, Rosetta-line,
P. O. Box: 22758, Egypt; gaber.abdellatif@alexu.edu.eg
M. M. Elnaggar: Department of Microbiology, Faculty of
Veterinary Medicine, Alexandria University, Edfina, Rosetta-line,
P. O. Box: 22758, Egypt; mahmoud.elnaggar@alexu.edu.eg
S. A. Khaliel: Department of Microbiology, Faculty of
Veterinary Medicine, Alexandria University, Edfina, Rosetta-line,
P. O. Box: 22758, Egypt; khaliel1@yahoo.com
A. E. Gamal-Eldin: Department of Microbiology, Faculty of
Veterinary Medicine, Alexandria University, Edfina, Rosetta-line,
P. O. Box: 22758, Egypt; dr_asmaa9518@yahoo.com
Received: 03-05-2014, Revised: 02-07-2014, Accepted: 10-07-2014,
Published online: 11-08-2014
Corresponding author: Gaber Abdellrazeq, email:
gaber.abdellatif@alexu.edu.eg
Abstract
Background: Johne's disease
(JD) caused by Mycobacterium avium subsp. paratuberculosis (MAP)
represents a real threat to the agriculture and dairy food
industries and believed to be a potential public health problem.
Signs of infection in ruminant include weight loss, diarrhea,
decreased milk production, and eventually death. The definition of
an infected animal based either on the presence of anti-MAP
antibodies, or positive bacterial culture. No treatment for the
disease exists and controlling the disease is difficult due to its
long latent period. JD is a worldwide problem and multiple studies
in many countries have been carried out to determine the
prevalence of MAP infections. Although some primary non intensive
studies confirm presence of JD in Egypt, the disease is currently
neglected by the official Egyptian veterinary agencies. There is
no official data, no national control program, and no used
vaccine.
Aim: This study aimed to evaluate three conventional
diagnostic methods for MAP under the Egyptian circumstances with a
general aim to determine the appropriate strategy to develop a JD
control program. These methods were pooled fecal culture, humoral
response and insertion sequence IS900 targets polymerase chain
reaction (IS900 PCR).
Materials and Methods: Fecal and serum samples (500 each)
were collected from Holstein-Friesian cattle and buffaloes housed
in five Egyptian governorates. Fecal samples were examined for MAP
on the basis of a strategic pooling procedure and performed on
Herrold's Egg Yolk Agar Medium (HEYM). Smears were prepared from
developed colonies and stained using a Ziehl-Neelsen (ZN)
technique. The identity of developed colonies was further
confirmed by PCR analysis of IS900 sequence. Sera from both
culture-positive and culture-negative animals were evaluated
individually for humoral response.
Results: Out of 50 pooled specimens, 34 (68%) fecal
cultures were positive for MAP. Serum positive samples of
culture-positive animals were 63/340 (18.5%). No serum positive
were detected in the samples from culture-negative animals. DNA
extractions from colonies of 34 culture-positive pooled samples
were tested with the PCR targeted to IS900 and revealed 18
isolates (52.94%) were positive.
Conclusion: The present study has succeeded in isolating
and identifying of MAP from Egyptian cattle and buffaloes in five
governorates. The results confirm the epizoology of JD in Egypt
and encourage the decision-makers to start creating a program to
control it. Pooling procedure for MAP culture is an efficient and
reliable method. However, the findings suggest the importance of
using both ELISA and fecal culture in MAP diagnosis and control.
Keywords: culture, ELISA, IS900 PCR, Johne's disease,
Mycobacterium avium subsp. paratuberculosis.
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