scholarly journals Detection of Mycobacterium avium Subspecies paratuberculosis in Milk and fecal Samples in Dairy Cattle by PCR and Nested-PCR

2010 ◽  
Vol 9 (24) ◽  
pp. 3055-3061 ◽  
Author(s):  
M. Tohidi Moghadam ◽  
S. Sarv ◽  
F. Moosakhani ◽  
A. Badiie
Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1605
Author(s):  
Annika Wichert ◽  
Esra Einax ◽  
Natalie Hahn ◽  
Anne Klassen ◽  
Karsten Donat

Within paratuberculosis control programs Mycobacterium avium subsp. paratuberculosis (MAP)-infected herds have to be detected with minimum effort but with sufficient reliability. We aimed to evaluate a combination of random sampling (RS) and pooling for the detection of MAP-infected herds, simulating repeated RS in imitated dairy herds (within-herd prevalence 1.0%, 2.0%, 4.3%). Each RS consisted of taking 80 out of 300 pretested fecal samples, and five or ten samples were repeatedly and randomly pooled. All pools containing at least one MAP-positive sample were analyzed by culture and real-time quantitative PCR (qPCR). The pool detection probability was 47.0% or 45.9% for pools of size 5 or 10 applying qPCR and slightly lower using culture. Combining these methods increased the pool detection probability. A positive association between bacterial density in pools and pool detection probability was identified by logistic regression. The herd-level detection probability ranged from 67.3% to 84.8% for pools of size 10 analyzed by both qPCR and culture. Pools of size 10 can be used without significant loss of sensitivity compared with pools of size 5. Analyzing randomly sampled and pooled fecal samples allows the detection of MAP-infected herds, but is not recommended for one-time testing in low prevalence herds.


Author(s):  
Nuzhat Hassan ◽  
Charanjit Singh Randhawa ◽  
Deepti Narang ◽  
Shoor Vir Singh ◽  
Shukriti Sharma ◽  
...  

Dairy cattle and buffaloes (n=101) suffering from chronic diarrhea (>3 weeks) with complete history of duration of diarrhea, systemic effects, body condition and faecal abnormalities were included in this study. Faecal samples, whole blood and rectal biopsies from selected animals were collected for the detection of Mycobacterium avium-subspecies paratuberculosis (MAP). Faecal smears were prepared and examined for the load of acid fast bacilli (AFB). Acid fast clumps were recorded in 40.6 per cent (41/101) of diarrheic animals and PCR confirmation was made byIS900 primers. MAP was detected in blood samples of four animals (4/41), while faecal samples were positive only in two of these 4 animals. All the four animals had persistent watery diarrhea and progressive loss in body weights and productivity. Rectal biopsies of two animals were positive by PCR. Remission of clinical signs was observed in these animals within 2-months. Biotyping of MAP-DNA from blood samples by IS1311 PCR-REA revealed restriction pattern of 67, 218 and 325 bp suggesting that MAP DNA belonged to ‘Bison type’, which is a major biotype infecting domestic livestock in India.


2020 ◽  
Vol 5 (6) ◽  
pp. 212-217
Author(s):  
S. Singh ◽  
◽  
I. P. Dhakal ◽  
U. M. Singh ◽  
B. Devkota ◽  
...  

Paratuberculosis caused by Mycobacterium avium subspecies paratuberculosis (MAP), is an economically important, endemic in many parts of the globe, and regarded as high prevalent disease of domestic and wild animals, especially ruminants, which is manifest as chronic granulomatous enteritis with decreased milk production, with serious cases resulting in progressive emaciation and death. Understanding the genetic variability of MAP, strains are important in diagnosis, epidemiological investigation, and therefore the formation of strategies for prevention and control of the disease. Thus, this study was designed to grasp the molecular characterization of MAP isolates of Nepal, as pioneer research of this area. Total of 46 MAP isolates obtained from cattle population of three different locations of dairy pocket areas of Chitwan, Nepal were typed using IS1311 polymerase chain reaction-restriction endonuclease analysis (PCR-REA) to research the MAP genotype of Nepal. The extracted DNA samples (n=46) were analyzed for the presence of MAP specific sequences (IS900) using PCR and DNA samples were further subjected to genotype differentiation using IS1311 PCR-REA and IS1311 L2 PCR-REA methods. All the DNA samples were positive for the entire three MAP specific sequences based PCRs. This study revealed that ‘Bison type’ strain is the single most prevalent MAP genotype circulating within the domestic cattle population of Nepal. IS1311 PCR-REA showed that MAP DNA samples of Nepal origin belonged to ‘Bison type’, whereas, IS1311 L2 PCR-REA method showed similarity with "Indian Bison type" and different restriction profiles of ‘Bison type’ genotype as compared to non-Indian strains. The study concludes that in Nepal, "Bison type" MAP stains was prevalent in all the MAP samples obtained from dairy cattle. These results have important epidemiological implications regarding control and prevention of paratuberculosis in Nepal.


2010 ◽  
Vol 93 (10) ◽  
pp. 4455-4470 ◽  
Author(s):  
C. Marcé ◽  
P. Ezanno ◽  
M.F. Weber ◽  
H. Seegers ◽  
D.U. Pfeiffer ◽  
...  

2018 ◽  
Vol 44 (04) ◽  
pp. 189-196
Author(s):  
Heng-Ching Lin ◽  
Wai How Chong ◽  
Han Hsiang Huang ◽  
Chi-Chung Chou ◽  
Yi-Lun Tsai ◽  
...  

Mycobacterium avium subspecies paratuberculosis (MAP) are Gram positive, aerobic, acid-fast, catalase positive bacteria. The Johne’s disease, caused by MAP, is a wasting disease found in all ruminants including cattle, sheep, goats, deer, camelids and wildlife ruminants. MAP has attracted hygienic attention due to the link between Crohn’s disease (CD) in humans and MAP presence in the gut of patients. The aims of this study are to investigate and monitor the serological prevalence and molecular identification rate of MAP in caprine feces and verify the MAP-negative goat farms in central-southern Taiwan. A total of 8486 blood samples were randomly collected between the years 2011 and 2015 from 321 caprine farms. The serum samples were assessed by commercial ELISA while 3015 fecal samples from 201 anti-MAP antibodies (MAP-Ab) negative herds were further molecularly examined by polymerase chain reaction (PCR) from year 2014 to 2015. The individual seroprevalence rate of caprine MAP in 2011, 2012, 2013, 2014 and 2015 was 0% (0/1032), 0% (0/429), 0% (0/1402), 0.14% (4/2917) and 0.07% (2/2706), respectively. Molecular identification rate of MAP in caprine fecal samples at MAP-Ab negative farms accounted for 0.92% (14/1515) and 0.93 (14/1500), respectively, in the years 2014 and 2015. Meanwhile, there was no association between the MAP seroprevalence and the sampling regions or years at farm or individual level. The seroprevalence data revealed in this study highlighted the rising prevalence of caprine MAP and the link of MAP to farmed ruminant species and its possible implications in hygienic aspects.


2008 ◽  
Vol 127 (3-4) ◽  
pp. 300-308 ◽  
Author(s):  
María C. Antognoli ◽  
Franklyn B. Garry ◽  
Heather L. Hirst ◽  
Jason E. Lombard ◽  
Michelle M. Dennis ◽  
...  

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