Molecular characterization of Mycobacterium avium subsp. paratuberculosis (MAP) in dairy cattle of Nepal

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.

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Culture phenotypes and molecular characterization of Mycobacterium avium subsp. paratuberculosis isolates from small ruminants

Research in Veterinary Science ◽

10.1016/j.rvsc.2013.03.010 ◽

2013 ◽

Vol 95 (1) ◽

pp. 49-53 ◽

Cited By ~ 10

Author(s):

Z. Dimareli-Malli ◽

K. Mazaraki ◽

K. Stevenson ◽

P. Tsakos ◽

A. Zdragas ◽

...

Keyword(s):

Molecular Characterization ◽

Mycobacterium Avium ◽

Small Ruminants ◽

Mycobacterium Avium Subsp ◽

Mycobacterium Avium Subsp Paratuberculosis

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Morphological and Molecular Characterization of a New Mycobacterium avium Subsp. paratuberculosis S-Type Strain Genotype in Goats

Frontiers in Veterinary Science ◽

10.3389/fvets.2019.00250 ◽

2019 ◽

Vol 6 ◽

Author(s):

Simone Scherrer ◽

Roger Stephan ◽

Jon Paulin Zumthor ◽

Anja Kipar ◽

Frauke Seehusen

Keyword(s):

Molecular Characterization ◽

Mycobacterium Avium ◽

Type Strain ◽

Mycobacterium Avium Subsp ◽

Mycobacterium Avium Subsp Paratuberculosis ◽

Morphological And Molecular Characterization

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Characterization of Mycobacterium avium subspecies paratuberculosis disseminated infection in dairy cattle and its association with antemortem test results

Veterinary Microbiology ◽

10.1016/j.vetmic.2007.08.017 ◽

2008 ◽

Vol 127 (3-4) ◽

pp. 300-308 ◽

Cited By ~ 39

Author(s):

María C. Antognoli ◽

Franklyn B. Garry ◽

Heather L. Hirst ◽

Jason E. Lombard ◽

Michelle M. Dennis ◽

...

Keyword(s):

Dairy Cattle ◽

Mycobacterium Avium ◽

Mycobacterium Avium Subspecies Paratuberculosis ◽

Test Results ◽

Disseminated Infection

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Serological and Molecular Characterization of Mycobacterium avium Subsp. paratuberculosis (MAP) from Sheep, Goats, Cattle and Camels in the Eastern Province, Saudi Arabia

Animals ◽

10.3390/ani11020323 ◽

2021 ◽

Vol 11 (2) ◽

pp. 323

Author(s):

Ibrahim Elsohaby ◽

Mahmoud Fayez ◽

Mohamed Alkafafy ◽

Mohamed Refaat ◽

Theeb Al-Marri ◽

...

Keyword(s):

Saudi Arabia ◽

Molecular Characterization ◽

Mycobacterium Avium ◽

Mycobacterium Avium Subsp ◽

Type I ◽

Serum Samples ◽

Eastern Province ◽

Fecal Samples ◽

Mycobacterium Avium Subsp Paratuberculosis

The objectives of the present study were to characterize Mycobacterium avium subsp. paratuberculosis (MAP) infection using serological and molecular tools and investigate the distribution and molecular characterization of MAP strains (cattle (C) and sheep (S) types) in sheep, goat, cattle, and camel herds in Eastern Province, Saudi Arabia. Serum and fecal samples were collected from all animals aged >2 years old in 31 herds (sheep = 8, goats = 6, cattle = 8 and camels = 9) from January to December 2019. Serum samples were tested by ELISA for the detection of MAP antibodies. Fecal samples were tested by PCR for the detection of MAP IS900 gene and the identification of MAP strains. MAP antibodies were detected in 19 (61.3%) herds. At the animal level, antibodies against MAP were detected in 43 (19.5%) sheep, 21 (17.1%) goats, 13 (19.7%) cattle and 22 (9.1%) camels. The IS900 gene of MAP was detected in 23 (74.2%) herds and was directly amplified from fecal samples of 59 (26.8%) sheep, 34 (27.6%) goats, 20 (30.3%) cattle and 36 (15.0%) camels. The S-type was the most prevalent MAP type identified in 15 herds, and all were identified as type-I, while the C-type was identified in only 8 herds. The IS900 sequences revealed genetic differences among the MAP isolates recovered from sheep, goats, cattle and camels. Results from the present study show that MAP was prevalent and confirm the distribution of different MAP strains in sheep, goat, cattle and camel herds in Eastern Province, Saudi Arabia.

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Molecular characterization of Mycobacterium avium subsp. paratuberculosis (MAP) isolates

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2017.v10i5.17310 ◽

2017 ◽

Vol 10 (5) ◽

pp. 393

Author(s):

Selvam Arjunan

Keyword(s):

Molecular Characterization ◽

Mycobacterium Avium ◽

Mycobacterium Avium Subsp ◽

Intermediate Type ◽

Chain Reaction ◽

Type Iii ◽

Polymerase Chain ◽

Mycobacterium Avium Subsp Paratuberculosis ◽

Digital Microfluidic

Objective: Johne’s disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is highly prevalent in domestic ruminants. In India, the exact prevalence of MAP genotypes still remains unknown limited, and the systematic disease control programs are also limited. This study was undertaken to study the molecular characterization of MAP isolates.Methods: About 22 MAP isolates were from cattle, sheep, and goat under gone the molecular characterization by three different methods (1) IS1311 polymerase chain reaction (PCR) with restriction enzyme analysis (REA), (2) GyrA and GyrB PCR with sequencing, and (3) digital microfluidic chip (DMC)-PCR. The study demonstrated that a) IS1311 PCR with REA (based on point mutations) identified all 22 MAP isolates as “intermediate type” irrespective of a host of origin and also belong to Indian Bison type. Molecular typing based on the gyrA and gyrB genes partial amplification and sequencing revealed that the MAP isolates exhibited more lineages toward the reference Type III, Intermediate strain.Results: The MAP isolate of sheep origin showed more lineages toward the sheep type than the isolates of cattle and goats. This variation may be due to host-pathogen interactions and adaptation to different hosts and environmental conditions in the nature.Conclusions: The DMC-PCR, which is based on sequence difference at 5` end of IS900 of MAP, differentiated rapidly all the isolates as sheep type. The application of DMC-PCR to differentiate sheep and Intermediate types is limited as the Intermediate type (Type III) and sheep type (Type I) are very closely related to each other and all the MAP isolates were confirmed as Intermediate or Type III by three different methods which are commonly present in India, Spain, and Iceland.Keywords: Johne’s disease, Mycobacterium avium subsp. paratuberculosis, Polymerase chain reaction, Digital microfluidic chip-polymerase chainreaction.

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