Identification of members of Mycobacterium avium species by Accu-Probes, serotyping, and single IS900, IS901, IS1245 and IS901-flanking region PCR with internal standards

Bartoš, Milan; Hložek, Pavel; Svastova, Petra; Dvorská, L.; Bull, Tim J.; Mátlová, L.; Parmova, I.; Kuhn, Isolde; Stubbs, Janine; Morávková, Monika; Kintr, Jaromir; Beran, V.; Melichárek, I.; Ocepek, M.; Pavlík, I.

doi:10.1016/j.mimet.2005.05.009

Cited by 78 publications

(65 citation statements)

References 45 publications

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“…This mix was submitted to the following PCR cycles: initial denaturation at 94 C for 5 min followed by 35 cycles of 94 C for 1 min, annealing at 60 C for 1 min, and extension at 72 C for 1 min with a final extension step of 72 C for 10 min. For IS1245 we followed protocols of Guerrero et al (1995) and Bartos et al (2006). Briefly, the same PCR mix was used, excluding the DMSO, with the following PCR cycles: 45 cycles of 94 C for 1 min, 65 C for 1 min and then 72 C for 1 min, with a final extension step of 72 C for 10 min.…”

Section: Bacterial Culture and Identificationmentioning

confidence: 99%

“…Polymerase chain reaction products were visualized by electrophoresis in 1% agarose gel with ethidium bromide and photographed under UV light (Alpha Imager, Alpha Innotech Corporation, San Leandro, California, USA). According to Huard et al (2003) and Bartos et al (2006), this set of genes allows for the identification of M. bovis, Mycobacterium caprae, other members of the Mycobacterium tuberculosis complex, members of the Mycobacterium avium complex, and other mycobacteria not belonging to these two complexes (Table 2).…”

Section: Bacterial Culture and Identificationmentioning

confidence: 99%

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Epidemiology of Mycobacterium Bovis Infection in Wild Boar (Sus Scrofa) From Portugal

Santos

Correia-Neves

Ghebremichael

et al. 2009

Journal of Wildlife Diseases

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ABSTRACT:Tuberculosis has been diagnosed in wild boar (Sus scrofa) in several European countries during the last decade; however, almost no information has been reported to date for Portugal. This study aimed to investigate tuberculosis in wild boar in Portugal through characterization of Mycobacterium bovis infection and identification of disease risk factors. Tissue samples were obtained from hunted wild boar during the 2005 and 2006 hunting seasons. Samples were inspected for gross lesions and processed for culture. Acid-fast bacterial isolates were identified by polymerase chain reaction and spoligotyping. Associations between tuberculosis in wild boar and several variables linked to wild ungulate diversity and relative abundance, livestock density, and cattle tuberculosis incidence were investigated. Mycobacterium bovis isolates were identified in 18 of 162 wild boars from three of eight study areas. Infection rates ranged from 6% (95% confidence interval [CI P95% ]51-21%) to 46% (CI P95% 527-67%) in the three infected study areas; females in our sample were at greater risk of being infected than males (odds ratio54.33; CI P95% 53.31-5.68). Spoligotyping grouped the M. bovis isolates in three clusters and one isolate was a novel spoligotype not previously reported in international databases. Detection of M. bovis was most consistently associated with variables linked to wild ungulate relative abundance, suggesting that these species, particularly the wild boar, might act as maintenance hosts in Portugal.

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Section: Bacterial Culture and Identificationmentioning

confidence: 99%

Section: Bacterial Culture and Identificationmentioning

confidence: 99%

Epidemiology of Mycobacterium Bovis Infection in Wild Boar (Sus Scrofa) From Portugal

Santos

Correia-Neves

Ghebremichael

et al. 2009

Journal of Wildlife Diseases

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“…avium (MAA) of serotypes 1, 2 and 3 and of genotype dnaJ+, IS901+ and IS1245+. The main sources of the causative agent of avian tuberculosis are infected domestic and wild birds and small rodents (Pavlik et al, 2000;Fischer et al, 2001;Mijs et al, 2002;Matlova et al, 2003;Bartos et al, 2006;Dvorska et al, 2007).…”

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confidence: 99%

Isolation of conditionally pathogenic mycobacteria from the environment of one pig farm and the effectiveness of preventive measures between 1997 and 2003

Pavlík¹,

Mátlová²,

Gilar³

et al. 2007

Vet. Med. - Czech

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392Tuberculosis in pigs causes high economic losses farmers from all over the world. The most consequential causative agents of tuberculosis in pigs at present are members of Mycobacterium tuberculosis (MTC) and M. avium (MAC) complexes. The losses are above all caused by restriction of animal transport from infected farms (with the exception of slaughterhouses), culling of animals positive during the tuberculin testing and the price of meat and organs from slaughtered infected animals (Alfredsen and Skjerve 1993;Dey and Parham, 1993;Morita et al., 1994;Balian et. al., 1997;Cvetnic et al., 1998; Positive skin responses were found in animals with antibodies against MAH (18; 23.7%), MAA (3; 4.0%) and MI (9; 11.8%) antigens. By serological examination of 17 sows with repeated dubious responses for tuberculin skin testing with avian tuberculin, no antibodies against MAA were detected; MAH antibodies and MI antibodies were found in eight and two animals, respectively. By post mortem examination of lymph nodes (ln) and organ samples from all 76 animals with responses to avian tuberculin, no tuberculous/tuberculoid lesions were detected. By culture examination of ln and organs from 13 animals, conditionally pathogenic mycobacteria (CPM) were isolated from only one animal (breeding boar): from mesenteric, pulmonary, hepatic ln and from spleen tissue samples. These isolates were identified as MAH and CPM by the PCR method and biochemically. By investigation of the external environment (205 samples), 33 (16.3%) CPM isolates were obtained: 13 MAH, eight M. fortuitum, one M. nonchromogenicum, one M. abscessus, one M. scrofulaceum and nine unidentified isolates, which were non-MAC according to the PCR examination. Non-specific responses obtained in the intravital tests (skin and serological tests) caused by CPM present in the environment substantially complicated diagnosis of avian tuberculosis. Based on these findings, animal hygiene measures have been adopted since 2002; resulted in a decrease of environmental contamination with CPM and a reduction in the number of animals giving positive responses to avian tuberculin.

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“…Sediment (200 ml) of each decontaminated sample was cultured on three slopes of different Herrold egg yolk media with Mycobactin J and incubated at 37 C for 12 mo (Machackova et al, 2004). Isolates of M. a. paratuberculosis were identified by IS900 polymerase chain reaction (PCR; Bartos et al, 2006). Standardized IS900 restriction fragment length polymorphism (RFLP) by using restriction endonucleases PstI and BstEII was used to further differentiate M. a. paratuberculosis isolates (Pavlik et al, 1999a).…”

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confidence: 99%

Detection of Mycobacterium avium subsp. paratuberculosis in Two Brown Bears in the Central European Carpathians

Kopecna

Ondrus²,

Literák

et al. 2006

Journal of Wildlife Diseases

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ABSTRACT:The incidence of mycobacterial infections was monitored in brown bears (Ursus arctos) in the National Park Low Tatras in the central European Carpathians in Slovakia. Tissue samples of 20 brown bears were examined microscopically and by culture for the presence of mycobacteria. Acid-fast rods were detected by Ziehl-Neelsen staining in a smear from the kidney of one brown bear, although the culture was negative for mycobacteria. Mycobacterium avium subsp. paratuberculosis, the causative agent of paratuberculosis in ruminants, was isolated from the intestinal mucosa of another two brown bears. The isolates were identified by polymerase chain reaction for the specific insertion sequence IS900. Using standardized IS900 restriction fragment length polymorphism (RFLP) analysis, the M. a. paratuberculosis isolates were classified as RFLP type B-C1, which also were detected in the infected cattle in surrounding area. This study describes the first isolation of M. a. paratuberculosis from a brown bear. Our results confirm that animal species other than ruminants can become infected with M. a. paratuberculosis and can act as potential vectors and/or reservoirs of the infection.

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Identification of members of Mycobacterium avium species by Accu-Probes, serotyping, and single IS900, IS901, IS1245 and IS901-flanking region PCR with internal standards

Cited by 78 publications

References 45 publications

Epidemiology of Mycobacterium Bovis Infection in Wild Boar (Sus Scrofa) From Portugal

Epidemiology of Mycobacterium Bovis Infection in Wild Boar (Sus Scrofa) From Portugal

Isolation of conditionally pathogenic mycobacteria from the environment of one pig farm and the effectiveness of preventive measures between 1997 and 2003

Detection of Mycobacterium avium subsp. paratuberculosis in Two Brown Bears in the Central European Carpathians

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