Detection of Mycobacterium avium subsp. paratuberculosis (MAP) from Subclinical Caprine Paratuberculosis Cases of Odisha

Paratuberculosis is caused by Mycobacterium avium subsp. Paratuberculosis (MAP) and is a chronic, intestinal tract infection in the ruminant sector globally. A total of 122 EDTA mixed blood samples, 121 serum and 16 pooled faecal samples were collected from farms of 4 different districts i.e. Nayagarh, Cuttack, Khordha and Angul and a blind review was conducted at the Animal Health Division, CIRG, Mathura. Microscopic examination of 16 pooled faecal samples revealed +2 reactivity to Acid-Fast Bacilli. All the serum samples were subjected to indirect ELISA. Out of them, 23 (19.01%), 85 (70.25%), shows strongly positive, positive, antibody titre respectively. EDTA blood samples of 23 ELISA-strongly positive were subjected to 413 bp IS900 PCR and 11 (9%) of them were found positive for Mycobacterium avium subsp. Paratuberculosis (MAP). MAP isolates were further subjected to genotyping using 608 bpIS1311 PCR and restriction endonuclease analysis (IS1311 PCR-REA) and 2 (1.64%) of them matched with “Indian Bison Type”. Genotyping of the isolates using IS1311 PCR-REA revealed that goat population of Odisha are primarily infected with “Indian Bison Type” strains.

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Faecal bacterial composition in dairy cows shedding Mycobacterium avium subsp. paratuberculosis in faeces in comparison with nonshedding cows

Canadian Journal of Microbiology ◽

10.1139/cjm-2015-0814 ◽

2016 ◽

Vol 62 (6) ◽

pp. 538-541 ◽

Cited By ~ 6

Author(s):

Marija Kaevska ◽

Petra Videnska ◽

Karel Sedlar ◽

Iva Bartejsova ◽

Alena Kralova ◽

...

Keyword(s):

Dairy Cows ◽

Mycobacterium Avium ◽

Mycobacterium Avium Subsp ◽

Rrna Gene ◽

Microbial Composition ◽

Bacterial Composition ◽

Bacterial Populations ◽

Faecal Samples ◽

Mycobacterium Avium Subsp Paratuberculosis ◽

The 16S Rrna Gene

The aim of this study was to determine possible differences in the faecal microbiota of dairy cows infected with Mycobacterium avium subsp. paratuberculosis (Johne’s disease) in comparison with noninfected cows from the same herds. Faecal samples from cows in 4 herds were tested for M. avium subsp. paratuberculosis by real-time PCR, and faecal bacterial populations were analysed by 454 pyrosequencing of the 16S rRNA gene. The most notable differences between shedding and nonshedding cows were an increase in the genus Psychrobacter and a decrease in the genera Oscillospira, Ruminococcus, and Bifidobacterium in cows infected with M. avium subsp. paratuberculosis. The present study is the first to report the faecal microbial composition in dairy cows infected with M. avium subsp. paratuberculosis.

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Detection of Mycobacterium avium subsp. paratuberculosis by several diagnostics techniques in clinical suspected sheep

Journal of the Hellenic Veterinary Medical Society ◽

10.12681/jhvms.15601 ◽

2018 ◽

Vol 68 (2) ◽

pp. 167

Author(s):

A. C. COELHO ◽

A. M. COELHO ◽

J. GARCÍA-DIEZ ◽

M. A. PIRES ◽

M. L. PINTO

Keyword(s):

Mycobacterium Avium ◽

Bacterial Culture ◽

Histopathological Examination ◽

Diagnostic Techniques ◽

Clinical Findings ◽

Mycobacterium Avium Subsp ◽

Clinical Symptomatology ◽

Faecal Samples ◽

Polymerase Chain ◽

Mycobacterium Avium Subsp Paratuberculosis

A total of thirty sheep with clinical symptomatology of paratuberculosis (Johne’s disease) were subjected to four diagnostic techniques: histopathological examination, bacteriological culture (in faeces and tissues), polymerase chain reaction (PCR) (in blood, tissue and faecal samples) and antibody responses (ELISA). Twenty-one (70.0%) animals showed histological lesions. Bacterial culture of both faeces and tissue revealed that 2 (6.7%) and 6 (20.0%) of the 30 sheep were infected, respectively. Mycobacterium avium subsp. 24 paratuberculosis (Map) was identified in 4 animals via PCR of faeces (13.3%), and in 19 (63.3%) by PCR in tissues. PCR in blood revealed 7 (23.3%) infected animals. Three (10.0%) animals showed antibodies against Map. A greater number of positive animals were detected by histopathological examination and PCR in tissues than by culture or ELISA. This study confirmed the clinical findings and results suggest that histopathology, PCR in tissues and in blood can improve the detection of Map in physically suspected animals and should be considered useful tools in the diagnosis of Map in suspected sheep.

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Efficiency of DNA Isolation Methods Based on Silica Columns and Magnetic Separation Tested for the Detection of Mycobacterium avium Subsp. Paratuberculosis in Milk and Faeces

Materials ◽

10.3390/ma13225112 ◽

2020 ◽

Vol 13 (22) ◽

pp. 5112

Author(s):

Marketa Husakova ◽

Petr Kralik ◽

Vladimir Babak ◽

Iva Slana

Keyword(s):

Magnetic Separation ◽

Mycobacterium Avium ◽

Magnetic Beads ◽

Dna Isolation ◽

Mycobacterium Avium Subsp ◽

Effective Control ◽

Silica Column ◽

Faecal Samples ◽

Isolation Methods ◽

Mycobacterium Avium Subsp Paratuberculosis

Timely and reliable detection of animals shedding Mycobacterium avium subsp. paratuberculosis (MAP) should help to effectively identify infected animals and limit infection transmission at early stages to ensure effective control of paratuberculosis. The aim of the study was to compare DNA extraction methods and evaluate isolation efficiency using milk and faecal samples artificially contaminated by MAP with a focus on modern instrumental automatic DNA isolation procedures based on magnetic separation. In parallel, an automatic and manual version of magnetic separation and two methods of faecal samples preparation were compared. Commercially available DNA isolation kits were evaluated, and the selected kits were used in a trial of automatic magnetic beads-based isolation and compared with the manual version of each kit. Detection of the single copy element F57 was performed by qPCR to quantify MAP and determine the isolation efficiency. The evaluated kits showed significant differences in DNA isolation efficiencies. The best results were observed with the silica column Blood and Tissue kit for milk and Zymo Research for faeces. The highest isolation efficiency for magnetic separation was achieved with MagMAX for both matrices. The magnetic separation and silica column isolation methods used in this study represent frequently used methods in mycobacterial diagnostics.

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Mycobacterium avium subsp. paratuberculosis ELISA Responses in Milk Samples from Vaccinated and Nonvaccinated Dairy Goat Herds in The Netherlands

Veterinary Sciences ◽

10.3390/vetsci6020058 ◽

2019 ◽

Vol 6 (2) ◽

pp. 58 ◽

Cited By ~ 2

Author(s):

Saskia Luttikholt ◽

Karianne Lievaart-Peterson ◽

Maaike Gonggrijp ◽

Marian Aalberts ◽

Gerdien van Schaik ◽

...

Keyword(s):

The Netherlands ◽

Mycobacterium Avium ◽

Relative Sensitivity ◽

Herd Size ◽

Mycobacterium Avium Subsp ◽

Dairy Goat ◽

Dairy Goats ◽

Milk Samples ◽

Goat Population ◽

Mycobacterium Avium Subsp Paratuberculosis

The aims of our study were to calculate the most appropriate cut-off value for milk samples in a serum-validated Mycobacterium avium subsp. paratuberculosis (MAP) ELISA and to analyze MAP ELISA responses in milk samples from vaccinated and nonvaccinated dairy goats in the Netherlands. Analyzed herds were representative for location and herd size of dairy goat herds in the Netherlands. A significantly higher proportion of the analyzed 49 herds were organic as compared with the total Dutch dairy goat population. First, the MAP ELISA was optimized using 992 paired serum and milk samples. At a cut-off of 25 S/P%, the relative sensitivity (Se) was 58.4% (n = 992, 95% CI: 48.8%−67.6%) and relative specificity (Sp) was 98.5% (n = 992, 95% CI: 97.5%−99.2%), as compared to serum ELISA results. The percentage of positively tested herds was 78.2% (n = 49, 95% CI: 63.4%−88.1%). The percentage of positive milk samples per herd (n = 22) was on average 4.6% (median, min, and max of 4.7%, 0.0%, and 10.7%, respectively). Average age of ELISA-positive (3.2 years) and -negative goats (3.2 years) was not different. Significantly more vaccinated goats tested positive (6.7%) as compared with nonvaccinated goats (1.1%). This study shows that a high number of vaccinated and nonvaccinated commercial dairy goat herds in the Netherlands have MAP-ELISA-positive goats.

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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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Detection of Mycobacterium avium subsp. paratuberculosis in Iranian patients with type 1 diabetes mellitus by PCR and ELISA

The Journal of Infection in Developing Countries ◽

10.3855/jidc.7473 ◽

2016 ◽

Vol 10 (08) ◽

pp. 857-862 ◽

Cited By ~ 4

Author(s):

Sonia Hesam Shariati ◽

Anoosha Alaei ◽

Rouhollah Keshavarz ◽

Nader Mosavari ◽

Ali Rabbani ◽

...

Keyword(s):

Diabetes Mellitus ◽

Type 1 Diabetes ◽

Type 1 Diabetes Mellitus ◽

Mycobacterium Avium ◽

Mycobacterium Avium Subsp ◽

Blood Samples ◽

Control Subjects ◽

Healthy Control ◽

Mycobacterium Avium Subsp Paratuberculosis

Introduction: Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis or Johne’s disease in ruminants. Its role in triggering autoimmunity, including type 1 diabetes mellitus (T1DM), has been reported in recent years. Due to the high contamination rate of MAP in Iran’s livestock and the increasing outbreak of T1DM, we investigated this association in a small group of patients with T1DM in Iran. Methodology: Blood samples of 29 T1DM patients and 29 healthy control subjects were tested through enzyme-linked immunosorbent assay (ELISA) to detect antibodies against MAP3865c and ZnT8 homologous epitopes and the presence of MAP DNA. Blood samples were also cultured in mycobacterial growth indicator tubes and Herrold's egg yolk medium containing mycobactin J. Results: The results of ELISA showed a significant difference between T1DM patients and healthy group. IS900 was also detected in 51.72% of T1DM patients but in none of the control group. None of the samples grew in culture media. Conclusions: Due to the presence of MAP DNA and antibodies against MAP peptides in a significant number of T1DM patients compared with healthy control subjects, we may consider MAP as a possible trigger of T1DM in Iran. This indicates that exposure to MAP occurred in the positive subjects. Identifying the sources of contamination and routes of MAP transmission to humans seems necessary to prevent and reduce the burden of MAP infection in Iran.

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