scholarly journals Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 20
Author(s):  
Luigi De Grossi ◽  
Davide Santori ◽  
Antonino Barone ◽  
Silvia Abbruzzese ◽  
Matteo Ricchi ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Liquid Culture ◽  
Culture Media ◽  
Small Ruminants ◽  
Culture Method ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Culture Methods ◽  
Specific Pcr ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is a chronic disease of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Since isolation of MAP type I (S) is rarely reported in Italy, our research was aimed at isolating, by an inexpensive liquid culture manual method, this type of MAP isolates. At first, we used an ELISA to point out to serologically positive samples from five flocks. Secondly, we used a fecal direct IS900-qPCR on the ELISA positive samples, in order to detect shedder animals. Feces from IS900-qPCR positive samples were inoculated in solid and liquid culture media. IS900-qPCR was further used to test the growth of MAP isolates in liquid medium, which were further confirmed by f57-qPCR and submitted to typing by specific PCR in order to identify the MAP type. Twenty-eight samples (24 fecal and four tissutal samples) were processed by culture methods, resulting in the isolation of six type I MAP field isolates. Notably, no isolates were recovered by solid media, underlining the utility of this liquid method. Few data about this type of MAP are currently available in Italy, and further analyses should be carried out in order to study the origin and epidemiology of type I strains circulating in Italy.

scholarly journals Effect of Egg Yolk on the Detection of Mycobacterium Avium Subsp. Paratuberculosis Using the ESP II Liquid Culture System

2005 ◽  
Vol 17 (6) ◽  
pp. 554-560 ◽  
Author(s):  
N. Beth Harris ◽  
Suelee Robbe-Austerman ◽  
Janet B. Payeur
Keyword(s):  
Mycobacterium Avium ◽  
Culture System ◽  
Liquid Culture ◽  
Egg Yolk ◽  
Mycobacterium Avium Subsp ◽  
Culture Methods ◽  
Solid Media ◽  
Liquid Culture System ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Time To Detection

Rapid diagnosis of paratuberculosis in infected cattle is important for the successful control of Johne disease within herds. Thus, improving culture methods for Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) will aid in the identification of asymptomatic animals. Egg yolk is a component of the media used for growing M. paratuberculosis, but its requirement as a supplement has not been reported. Using the ESP II liquid culture system, 2 different sources and 5 concentrations (3.3%, 1.6%, 0.8%, 0.4%, and 0%) of egg yolk were analyzed. Egg yolk source did not affect either recovery rate or time to detection, but both parameters were significantly improved when the 3.3% egg yolk concentrations (final volume) were used over media containing no egg yolk. This study also assessed the recovery of M. paratuberculosis from fecal samples that were cultured multiple times using Herrold egg yolk agar (HEY). Specimens containing greater than 70 cfu/g feces could routinely be identified as positive for M. paratuberculosis after only 1 culture attempt, whereas specimens with fewer bacteria were only intermittently positive, even after 5 replicate cultures. Therefore, this study indicates that the sensitivity of the Trek Diagnostic ESP II liquid culture system for M. paratuberculosis is affected by egg yolk concentration and that single culture attempts using HEY solid media may not identify specimens containing low numbers of bacteria.

scholarly journals Polymorphisms in gyrA and gyrB Genes among Mycobacterium avium subsp. paratuberculosis Type I, II, and III Isolates

2007 ◽  
Vol 45 (10) ◽  
pp. 3439-3442 ◽  
Author(s):  
E. Castellanos ◽  
A. Aranaz ◽  
B. Romero ◽  
L. de Juan ◽  
J. Alvarez ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Mycobacterium Avium Subsp Paratuberculosis

scholarly journals Association of Mycobacterium avium subsp. paratuberculosis and SLC11A1 polymorphisms in Sardinian multiple sclerosis patients

2013 ◽  
Vol 7 (03) ◽  
pp. 203-207 ◽  
Author(s):  
Davide Cossu ◽  
Speranza Masala ◽  
Eleonora Cocco ◽  
Daniela Paccagnini ◽  
Stefania Tranquilli ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Autoimmune Disease ◽  
Mycobacterium Avium ◽  
Humoral Response ◽  
Mycobacterium Avium Subsp ◽  
Nucleotide Polymorphisms ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Multiple Sclerosis Patients

Introduction: Recent findings propose that Mycobacterium avium subsp. paratuberculosis (MAP) infection could act as risk factor in favoring multiple sclerosis (MS) progression. SLC11A1 is a gene associated with mycobacterial survival in the host and it may be involved in the induction and maintenance of autoimmune disease. Methodology: In this preliminary study, 100 MS patients and 100 healthy controls (HCs) from Sardinia were enrolled. Eight single nucleotide polymorphisms (SNPs) in the SLC11A gene were searched by PCR RFLP-genotyping. IS900 specie specific PCR was undertaken to search for MAP presence. Indirect ELISA was performed to asses if MS patients displayed a stronger humoral response against MAP2694 protein compared to the HCs. Results: Only rs2276631 SNP was associated with MS. MAP DNA was detected in 23 out of 100 MS patients (23%) and in 7 out of 100 HCs (7%). A strong humoral response against MAP2694 protein was detected in 36% of MS patients and only in 3% of HCs. A correlation between ELISA sero-positivity and the rs2276631 SNP was also found. Conclusion:  Our preliminary results suggest that the Sardinian population might be prone to develop autoimmune disease due to polymorphisms in immunomodulating the SLC11A1 gene, which is important in the immune response against intracellular bacteria such as MAP.

Comparison of PCR Prescreening to Two Cultivation Procedures with PCR Confirmation for Detection of Mycobacterium avium subsp. paratuberculosis in U.S. Department of Agriculture Fecal Check Test Samples

2004 ◽  
Vol 67 (10) ◽  
pp. 2310-2314 ◽  
Author(s):  
JAY L. E. ELLINGSON ◽  
JEFFREY J. KOZICZKOWSKI ◽  
JENNIFER L. ANDERSON
Keyword(s):  
Mycobacterium Avium ◽  
Culture Medium ◽  
Mycobacterium Avium Subsp ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Culture Methods ◽  
Department Of Agriculture ◽  
Systems Research ◽  
Specificity And Sensitivity ◽  
Mycobacterium Avium Subsp Paratuberculosis

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent in Johne's disease in cattle and causes diarrhea, decreased milk production, emaciation, and frequently death. The ability to detect MAP rapidly and accurately is an integral part of herd management. However, detection of this bacterium is complicated due to its slow division time and its ability to enter dormancy. Culture methods are considered the “gold standard,” but they have their limitations. Many enzyme-linked immunosorbent assay methods and conventional PCR methods have been used as diagnostic tools. The present study compares the results of a PCR prescreen to two culture methods of detection paired with confirmatory PCR to determine the most accurate, rapid, and sensitive method using U.S. Department of Agriculture (USDA) fecal check samples. This study involving two laboratories (Marshfield Clinic Laboratories, using solid culture medium [Herrold's egg yolk agar], and TREK Diagnostic Systems Research and Development, using liquid culture medium [ESP Culture System II]) showed that the PCR prescreening method used in this study lacked specificity and sensitivity as a stand-alone test in fecal samples. However, the combination of liquid enrichment culture using the ESP II system, and PCR confirmation with the hspX primer set, was not only 100% sensitive and specific but also correlated with viable MAP and USDA culture results.

Detection of mycobacterium avium subsp. paratuberculosis in cheeses from small ruminants in Tuscany

2016 ◽  
Vol 217 ◽  
pp. 195-199 ◽  
Author(s):  
Alessia Galiero ◽  
Filippo Fratini ◽  
Antonia Mataragka ◽  
Barbara Turchi ◽  
Roberta Nuvoloni ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Small Ruminants ◽  
Mycobacterium Avium Subsp ◽  
Mycobacterium Avium Subsp Paratuberculosis

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

Animals ◽  
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.

Sign in / Sign up

Export Citation Format

Share Document