Strategy for the detection and differentiation of Mycobacterium avium species in isolates and heavily infected tissues

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The objectives of this study were to investigate Mycobacterium avium subsp. paratuberculosis (MAP) distribution in various tissues, in faeces and in milk samples from 131 animals originating from one imported Jersey cattle herd from Denmark. MAP was detected by culture in 37.4% animals. Massive MAP growth was most often observed in the small intestines (48 animals). The lowest levels of MAP were found in spleen and mammary gland samples. MAP was detected in the faeces of 8.4% animals; however, milk samples were MAP negative by culture. The highest prevalence of MAP infection (42.9%) was in age Group B (1.6 to 3 years) and the lowest (6.1%) in group D (more than 8 years). Seven positive calves were detected in the youngest age group; one of them was less than one month of age, which could imply an intrauterine infection. MAP shedding in faeces by a five-month-old calf was also confirmed. The IS900 RFLP (restriction fragment length polymorphism) type B-C1 was identified in all animals.

Section: Methodsmentioning

confidence: 99%

Distribution of Mycobacterium avium subsp.paratuberculosis in tissues of naturally infected cattle as affected by age

Hasoňová¹,

Trcka²,

Babák³

et al. 2009

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“…The cultures were checked after one week to rule out fast growing mycobacterial species or possible contamination and then every two weeks for the following two months. DNA was extracted from single colonies of the isolates as described by Moravkova et al (2008). Further identification was done by conventional PCR for the presence of specific mycobacterial DNA and MAC classification (Wilton and Cousins, 1992); and for presence of specific insertion sequences for subspecies identification .…”

Section: Mycobacterial Culture and Identification Of Isolatesmentioning

confidence: 99%

“…This method was determined to not be specific enough for the identification of MAA, as all the subspecies of M. avium species have 100% homology in their 16S rRNA gene sequences and are therefore not distinguishable from one another. With the discovery of insertion elements in the mycobacterial genome, the determination of M. avium subspecies has since became more precise (Shamputa et al, 2004;Moravkova et al, 2008). MAA is characterised by the presence of 2 to 17 copies of the IS901 insertion sequence (Dvorska et al, 2003;Inglis et al, 2003) and a single copy of IS1245 (Johansen et al, 2007).…”

mentioning

confidence: 99%

Examination of Mycobacterium avium subsp. avium distribution in naturally infected hens by culture and triplex quantitative real time PCR

Kaevska¹,

Slaná²,

Králík³

et al. 2010

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Mycobacterium avium subsp. avium (MAA) is the etiologic agent of avian tuberculosis, a chronic contagious disease described in a wide variety of domestic and wild bird species. The aims of this study were to assess the advantages of triplex quantitative real time PCR (qPCR) in comparison with culture testing for distribution of MAA in the organs of hens displaying varying degrees of clinical symptoms of the disease. From one small flock of ten hens and one cock with a history of weight loss, 98 tissue samples were examined in total. Pathological lesions were observed in six hens from which two were clinically ill. A total of 12 samples were positive by culture and 16 were positive by IS901 and IS1245 qPCR, confirming MAA infection. In conclusion, qPCR was a faster and more reliable alternative method in comparison with conventional culture analysis. Due to the detection of MAA in the muscle tissue of one hen, consumption of under cooked meat originating from infected fowl could pose a threat to immunosuppressed individuals.

“…Both tissue samples were cultured and yielded coats of colony forming units. The isolates were confirmed by conventional multiplex PCR as previously described (Moravkova et al, 2008).…”

Section: Case Descriptionmentioning

confidence: 99%

Avian tuberculosis in a captured Ruppell's griffon vulture (Gyps ruppellii): a case report

Škorič¹,

Fictum²,

Frgelecová³

et al. 2010

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Avian tuberculosis was diagnosed in a captured female Ruppell's griffon vulture (Gyps ruppellii) with granulomatous splenitis and hepatitis. At necropsy, whitish to yellow nodules of various sizes were found in the spleen and liver, and fibrinous coelomitis was present in the body cavity. Histopathologically, the granulomas appeared to be typical of avian tuberculosis. In some granulomas, necrotic centres surrounded by a variable layer of palisading epithelioid macrophages and multinucleated giant cells with variable admixture of lymphocytes and plasma cells were present. Signs of mineralization in granulomas were not observed. Using Ziehl-Neelsen staining the presence of acid-fast bacilli was demonstrated in organs affected by granulomatous inflammation. A diagnosis of Mycobacterium avium subsp. avium infection was confirmed by culture and quantitative Real-Time PCR examination for the presence of specific insertion sequences for avian tuberculosis (IS901 and IS1245) in the liver and spleen.